Pluronic-coated dye composition having improved stability and color retention and use thereof

Abstract

The present invention relates to a pluronic-coated dye composition having improved stability and color retention and use thereof. More specifically, the present invention relates to: a dye composition having improved stability and color retention, comprising dye nanoparticles and a pluronic polymer coated on the surface of the dye nanoparticles; and use of the dye composition in cosmetic compositions, ink compositions, paint compositions, resin compositions, food compositions, and the like.

Description

Pluronic coating dye composition having improved stability and color retention and use thereof

[0001] The present invention relates to a pluronic coating dye composition having improved stability and color retention and its use. More specifically, the present invention relates to a dye composition having improved stability and color retention comprising dye nanoparticles and a pluronic polymer coated on the surface of the dye nanoparticles, and to the use of the dye composition in cosmetic compositions, ink compositions, paint compositions, resin compositions, food compositions, and the like.

[0002] A tattoo is a method of creating letters or designs by incising the skin or subcutaneous tissue with a dye composition and then coloring it. Semi-permanent makeup is also available for eyebrows, eyeliner, lips, and hairlines. Recently, tattoos have been utilized and recognized as a way to compensate for physical weaknesses or to express individuality. Since tattoos inflict damage to the skin and color the skin with a dye composition, ensuring the safety of the dye is crucial. However, caution is required as heavy metals such as lead and nickel, which can cause cancer and allergies, are consistently detected in existing dyes. Furthermore, there have been numerous cases of pain, blisters, swelling, and mild inflammation after tattooing on areas such as the eyebrows, and cases of dye entering the eyes after eyeliner, causing pain and visual impairment.

[0003]

[0004] Prior art literature

[0005] Patent Document 1. Republic of Korea Patent Gazette No. 10-2451153

[0006] The present inventors have made extensive efforts to develop a dye having excellent stability and biocompatibility while also having improved color retention, and as a result, have confirmed that when dye nanoparticles are coated with a pluronic polymer, not only is dispersibility and storage stability excellent, but also biocompatibility is excellent and color retention ability is improved without being affected by oxidative stress, and thus have completed the present invention.

[0007]

[0008] However, the problems that the present invention seeks to solve are not limited to the problems mentioned above, and other problems not mentioned will be clearly understood by those skilled in the art from the description below.

[0009] The present invention seeks to provide a pluronic coating dye composition having improved stability and color retention and uses of the dye composition, for example, uses in cosmetic compositions, ink compositions, paint compositions, resin compositions or food compositions.

[0010]

[0011] The present invention

[0012] dye nanoparticles, and

[0013] The present invention relates to a dye composition having improved stability and color retention, comprising pluronics coated on the surface of the dye nanoparticles.

[0014] In the present invention, the dye nanoparticles may include edible charcoal (soot), brown amber (BA), gold shrine (GS), red amber (RA), silver shrine (SS), orange amber (OA), gold lustre (GL), light gold (LG), or a combination thereof.

[0015]

[0016] In one exemplary embodiment, the weight ratio of the edible charcoal nanoparticles and Pluronic may be 1:1 to 100. The Pluronic-coated edible charcoal nanoparticles may include edible charcoal and Pluronic PF127 in a weight ratio of 1:20.

[0017]

[0018] In another exemplary embodiment, the weight ratio of the brown amber (BA), gold shrine (GS), red amber (RA), silver shrine (SS), orange amber (OA), gold lustre (GL) or light gold (LG) and the Pluronic polymer may be 1:1 to 40. The dye composition may include brown amber (BA) and Pluronic F127 in a weight ratio of 1:20.

[0019] In the present invention, the dye composition can be used on hair, skin, eyes, eyebrows or eyelashes.

[0020] In the present invention, the dye composition can be used in a cosmetic composition, an ink composition, a paint composition, a resin composition, or a food composition.

[0021] The dye composition comprising the dye nanoparticles according to the present invention and the pluronic polymer coated on the surface of the dye nanoparticles not only has excellent dispersibility and storage stability, but also has excellent biocompatibility, is not affected by oxidative stress, and has significantly improved color retention ability.

[0022]

[0023] Meanwhile, the scope of the present invention is not limited by the effects described above.

[0024] Figure 1 shows (A) photographs, (B) UV-Vis spectra, (C) sizes, and (D) dispersion values ​​of Soot and Soot / Plu NPs.

[0025] Figure 2 shows the results of the dispersion stability evaluation of Soot and Soot / Plu NP, showing the UV-Vis spectra of Soot and Soot / Plu NP in DIW after storage at room temperature (25 ℃) for 2 weeks.

[0026] Figure 3 shows (A) the size and (B) dispersion values ​​of Soot and Soot / Plu NPs before and after lyophilization (FD).

[0027] Figure 4 shows the size change of Soot / Plu NPs after 4 weeks of storage in (A) DIW and (B) PBS at 37°C, and the dispersion change of Soot / Plu NPs after 4 weeks of storage in (C) DIW and (D) PBS at 37°C.

[0028] Figure 5 shows (A) the photograph, (B) the UV-Vis spectra, (C) the size, and (D) the dispersion values ​​of Soot / PF127 NP at different ratios.

[0029] Figure 6 shows the results of the dispersion stability evaluation of Soot / PF127 NP by ratio, showing the UV-Vis spectra of Soot / PF127 NP by ratio after storage in DIW at room temperature (25 ℃) for 2 weeks.

[0030] Figure 7 shows the (A) size and (B) dispersion values ​​of nanoparticles after freeze-drying (FD) and redispersion in DIW and PBS at different Soot / PF127 NP ratios.

[0031] Figure 8 shows the change in size after 4 weeks of storage in (A) DIW and (B) PBS at 37°C at different Soot / PF127 NP ratios, and the change in dispersion value after 4 weeks of storage in (C) DIW and (D) PBS at 37°C at different Soot / PF127 NP ratios.

[0032] Figure 9 shows the cytotoxicity results according to the concentration of Soot / PF127 NP (1:20).

[0033] Figure 10 shows cell morphology results according to the concentration of Soot / PF127 NP (1:20); (i) negative control, (ii) positive control, (iii) 0.1 mg / mL, and (iv) 1 mg / mL Soot / PF127 NP (1:20).

[0034] Figure 11 shows the results of oxidative stress induced by reactive oxygen radicals of Soot / PF127 NP (1:20).

[0035] Figure 12 shows the results of the 8-week tattoo retention ability of Soot / PF127 NP (1:20); the results of administration of the original solution (left) and a 1 / 2 dilution (right).

[0036] Figure 13 shows the H&E staining analysis results of (A) the original solution and (B) the 1 / 2 dilution after 4 weeks of Soot / PF127 NP (1:20) treatment, and (C) the original solution and (D) the 1 / 2 dilution after 8 weeks.

[0037] Figure 14 shows photographs and UV-Vis spectra of (a) brown amber (BA), (b) gold shrine (GS), (c) red amber (RA), and (d) silver shrine coated with PF127 or PVP (10 kDa).

[0038] Figure 15 shows (a) the size and (b) dispersion values ​​of four cosmetic dyes coated with PF127 or PVP (10 kDa).

[0039] Figure 16 shows (a) a photograph, (b) UV-Vis spectra, (c) size, and (d) dispersion values ​​of BA / PF127 according to the PF127 coating ratio.

[0040] Figure 17 shows (a) the size and (b) the dispersion values ​​of BA / PF127 after 4 weeks of storage at 37°C in DIW according to the PF127 coating ratio.

[0041] Figure 18 shows the cytotoxicity results according to the concentration of BA / PF127 (1:20).

[0042] Figure 19 shows the cell morphology results according to the concentration of BA / PF127 (1:20): (i) negative control, (ii) positive control, (iii) 0.1 mg / mL, and (iv) 1 mg / mL BA / PF127 (1:20) (Scale bar = 100 μm).

[0043] Figure 20 shows the results of oxidative stress induced by reactive oxygen radicals of BA / PF127 (1:20).

[0044] Figure 21 shows the results (a) immediately after administration and (b) 4 weeks after administration of BA / PF127 (1:20).

[0045] Hereinafter, a pluronic coating dye composition having improved stability and color retention according to specific embodiments of the invention and its uses will be described in detail. However, this is presented as one example of the invention, and the scope of the invention is not limited thereby, and it will be apparent to those skilled in the art that various modifications to the embodiments are possible within the scope of the invention. Unless otherwise specified, throughout this specification, "include" or "containing" refers to including a certain component (or component) without any particular limitation, and cannot be interpreted as excluding the addition of other components (or components).

[0046] The term "pluronic polymer" as used herein refers to an amphiphilic polymer having a structure of polyethylene oxide (PEO)-polypropylene oxide (PPO)-polyethylene oxide (PEO), which has several advantages such as a chemically cross-linked stable structure, in vitro and in vivo stability, ease and efficiency of protein loading, and non-cytotoxicity, as well as temperature sensitivity.

[0047]

[0048] 1. Pluronic coating dye composition

[0049] The present invention

[0050] dye nanoparticles, and

[0051] It is an object of the present invention to provide a dye composition having improved stability and color retention, comprising a pluronic polymer coated on the surface of the dye nanoparticles.

[0052] In the dye composition according to the present invention, the dye nanoparticles may include edible charcoal (soot), brown amber (BA), gold shrine (GS), red amber (RA), silver shrine (SS), orange amber (OA), gold lustre (GL), light gold (LG), or a combination thereof.

[0053] In the dye composition according to the present invention, the pluronic may include PF68, PF127, PP123, or a combination thereof.

[0054]

[0055] In one exemplary embodiment, when the dye nanoparticles are edible charcoal, the weight ratio of the edible charcoal nanoparticles and Pluronic may be 1:1 to 100, preferably 1:2 to 50, and most preferably 1:20. The edible charcoal and Pluronic PF127 may be included in a weight ratio of 1:20. The diameter of the Pluronic-coated edible charcoal nanoparticles may be 1 to 500 nm, preferably 1 to 400 nm, and most preferably 1 to 300 nm.

[0056]

[0057] In another exemplary embodiment, when the dye nanoparticles are brown amber (BA), gold shrine (GS), red amber (RA), silver shrine (SS), orange amber (OA), gold lustre (GL), or light gold (LG), the weight ratio of the dye nanoparticles to the pluronic polymer may be 1:1 to 50, preferably 1:5 to 40, and most preferably 1:20. The pluronic polymer may be Pluronic F127. The dye composition may include brown amber (BA) and pluronic polymer F127 in a weight ratio of 1:20. The diameter of the dye composition may be 1 to 800 nm, preferably 1 to 600 nm, and most preferably 1 to 500 nm.

[0058] In the dye composition according to the present invention, the dye composition can be used on hair, skin, eyes, eyebrows or eyelashes.

[0059]

[0060] 2. Use of Pluronic Coating Dye Composition

[0061] The present invention seeks to provide the use of the above 1. Pluronic coating dye composition in a cosmetic composition, ink composition, paint composition, resin composition, food composition, etc.

[0062]

[0063] (1) Cosmetic composition

[0064] In the use of the dye composition according to the present invention, the cosmetic composition is an eye shadow, an eyebrow, an eye liner, a mascara, an eyebrow mascara, an eyebrow quick tattoo, an aegyo-sal liner, a shade and shadow, a shading, a contouring, a concealer, a powder, a pact, a foundation, a base, a highlighter, a lipstick, a lip tint, a tint, a lip balm, a tinted lip balm, a lip gloss, a lip liner, a lotion, a toner, a serum, an eyelash serum, a mask pack, a sunscreen, a blusher, a plumper, an ampoule, an essence, a hair tint, a hair mist, a hair spray, a cream blusher, a lip serine, a sun cushion, a sun spray, a tone-up cream, a primer, a blur, a glitter, a lip and cheek, a fixer, a balm, a modeling pack, a patch, a skin pad, a cleansing balm, a spot gel, a black pigment, a black pigment spray, a shampoo, a rinse, It can be formulated as a treatment, hair mask, hair wax, hair gel, hairline cover, gray hair cover, hair dye, dye shampoo, toothpaste, bath salt, cleansing foam, cleansing oil, cleansing gel, cleansing water, cleansing mask, soap, shaving cream, shaving gel, massage cream, deodorant, body wash, scrub, cosmetic cotton swab, or cosmetic adhesive.

[0065] (2) Ink composition

[0066] In the use of the dye composition according to the present invention, the ink composition can be used for corneal tattoo, nipple tattoo, areola tattoo, scar tattoo, radiotherapy marker, eyebrow tattoo, eyeliner tattoo, lip tattoo, aegyo-sal tattoo, nail tattoo, tooth tattoo, scalp tattoo, scalp black tattoo, sideburn tattoo, beard tattoo, pubic tattoo, or henna tattoo.

[0067] In the use of the dye composition according to the present invention, the ink composition can be used as pen ink, fountain pen ink, printer ink, toner cartridge or art paint.

[0068] In the use of the dye composition according to the present invention, the ink composition can be used to print packaging packages, wrapping paper, wallpaper, fibers, cosmetic boards, textiles, various films or labels.

[0069]

[0070] (3) Paint composition

[0071] In the use of the dye composition according to the present invention, it can be used in ceramic paints, insecticides and high-efficiency spray fertilizers, ship paints, exteriors of automobiles, motorcycles and bicycles, building materials, roof tiles, furniture, household goods, containers, office supplies or sporting goods.

[0072]

[0073] (4) Resin composition

[0074] In the use of the dye composition according to the present invention, the resin composition can be used in containers for cosmetics, food containers, wall coverings, flooring, crafts, housings for home appliances, accessories, stationery, toys, bathtubs, bath supplies, shoes, sporting goods, wrist guards or toiletries.

[0075]

[0076] (5) Food composition

[0077] In the use of the dye composition according to the present invention, it can be used as a colorant for processed foods such as beverages, desserts, seasonings, spices, natural food coloring agents, animal feed, preservatives, capsules of health functional foods, confectionery, candy, breads, rice cakes, chocolates, beverages, alcoholic beverages, jellies, cereals, sugars, ice creams, seasonings, dressings, sauces, seasoned foods, pickles, agricultural and marine products, starch products, vegetable creams, ready-to-eat foods, grain products, edible oil products, sugar products, processed paper products, and processed dairy products.

[0078] (6) Other purposes

[0079] In the use of the dye composition according to the present invention, in addition to the above-mentioned use, the dye composition can be used in rubber, antistatic agents, semiconductive or conductive materials, papermaking, adsorption and deodorization, horticulture and agriculture, energy storage materials, directional carbon materials, abrasives, absorbent implants, or fire extinguishing agents as medicines.

[0080] In the use of the dye composition according to the present invention, the rubber can be used in automobile, aircraft or industrial tires, rubber seals, or packing.

[0081] In the use of the dye composition according to the present invention, the antistatic agent can be used in housings, cassettes, mechanical parts of electrical appliances, vehicle fuel tanks, hoses for chemical solvents, conveyor belts, V-belts, safety shoes, rollers for copiers, explosion-proof containers, integrated circuit packages (IC packages), storage warehouses, trays, containers, film canisters, magnetic tapes for video or audio, or backings (backings) of carpets in computer rooms.

[0082] In the use of the dye composition according to the present invention, the semiconductive and conductive materials can be used in a semiconductive compound, a cross-linked polyethylene cable, an ignition cable, a communication cable, a next-generation storage battery (Zn-Br type and Zn-Cl type), a ferrite magnet, a magnetic storage device (Hard Disk Drives-HDDs), magnetic particles of a magnetic cassette tape, a discharge resistor, a conductive element, an electric filter, or an electrostatic dissipation (ESD) material.

[0083] In the use of the dye composition according to the present invention, the adsorption and deodorization can be used as an activated carbon filter and mask, an air purifier and filter, an automobile air purifier, a deodorizer for shoes, an air freshener and a room freshener, a refrigerator deodorizer, a clothing and shoe storage box, a toilet, a deodorizer for baby diapers, and a moisture remover.

[0084] In the use of the dye composition according to the present invention, it can be used in the horticultural and agricultural fields as a biodeodorizer, biofertilizer, seed coating, and soil conditioner.

[0085] In the use of the dye composition according to the present invention, the energy storage material can be used in ultra-high capacity batteries, energy storage systems, automobiles and transportation vehicles, power grid stabilization, and mobile power supply devices.

[0086] In the use of the dye composition according to the present invention, the directional carbon material can be used in carbon fibers, carbon composites, electrically conductive and thermally conductive polymers (electronic devices, biosensors, LED lighting, automobile parts), deep-sea underwater exploration equipment, and military and aviation radar penetrators.

[0087] In the use of the dye composition according to the present invention, the dye composition can be used as a lens care solution, a multipurpose solution, or a wetting agent in the field of over-the-counter drugs.

[0088] In the use of the dye composition according to the present invention, the dye composition can be used as an auxiliary therapeutic agent or an adsorbent in the field of general pharmaceuticals.

[0089] In the use of the dye composition according to the present invention, the dye composition can also be used as a filtering aid for filtration, deodorization, decolorization, purification, etc. in food manufacturing or processing.

[0090] Below, various examples are presented to aid understanding of the invention. These examples are provided solely to facilitate understanding of the invention and are not intended to limit the scope of protection of the invention.

[0091]

[0092] <Example>

[0093] Example 1

[0094] 1-1. Preparation and Characterization of Five Types of Pluronic-Coated Edible Charcoal Nanoparticles (Soot / Plu NP)

[0095] (1) Experimental method

[0096] To develop a semi-permanent cosmetic dye with excellent biocompatibility and stability, Pluronic-coated edible charcoal nanoparticles (Soot / Plu NP) were prepared. First, to compare the characteristics of Soot / Plu NP by Pluronic type, five types of Pluronic (PF68, PF127, PL35, PP123, PL81) were prepared in 1 mL of deionized water (DIW) at a concentration of 25 mg / mL. The Pluronic solution was added to edible charcoal (soot; 5 mg), transferred to a 20 mL vial, and reacted with magnetic stirring at 300 rpm for 30 minutes. Then, 4 mL of DIW was added, and the reaction was continued at 400 rpm for an additional 30 minutes. The resulting Soot / Plu NPs were homogenized for 1 h using an ultrasonic homogenizer (amplitude = 20%, 5 s on / 15 s off), centrifuged (2000 rpm, 10 min) to remove uncoated edible charcoal, and then freeze-dried for 3 days. The Soot / Plu NPs thus manufactured were named Soot / PF68 NP, Soot / PF127 NP, Soot / PL35 NP, Soot / PP123 NP, and Soot / PL81 NP according to the type of coated Pluronic. Edible charcoal nanoparticles were also prepared using the same method without polymer coating. The morphology of Soot and Soot / Plu NPs was photographed, and their absorbance was analyzed using UV-Vis spectroscopy. The size and dispersion values ​​of the nanoparticles were measured using a Zetasizer (ELSZ-2000, Otsuka). In order to evaluate the dispersion stability of Soot / Plu NPs, for which the characteristics were analyzed in this way, the UV-Vis spectra of the dispersed nanoparticles were measured while storing them in DIW at room temperature (25 ℃) for 2 weeks. In addition, to evaluate the freeze-drying stability, the size and dispersion values ​​of the nanoparticles were analyzed before freeze-drying of soot and Soot / Plu NPs and after redispersion in DIW or PBS.The in vivo stability of nanoparticles was measured using a zetasizer device to measure changes in size and dispersion values ​​in DIW or PBS at 37°C for 4 weeks.

[0097] (2) Results

[0098] Characterization: As can be seen from Figure 1, both soot and Soot / Plu NPs were prepared as dark black solutions after ultrasonic homogenization (Figure 1A). At this time, Soot / PL81 NP was excluded from subsequent characterization due to poor stability after manufacturing, which caused partial precipitation (▲). Soot also had poor dispersion stability, and when the yield of soot contained in the nanoparticles was confirmed through the intensity of the unique UV-Vis absorption peak (λ = 260 nm) of soot, the absorbance was very low. However, after pluronic coating, the dispersibility of edible charcoal was improved, and the absorbance was also measured to be high (Fig. 1B). Among them, the yield was confirmed to be the highest when coated with PF127, PP123, or PL35. In addition, the size of soot before coating was approximately 241 nm, but after coating with PF68, PF127, and PP123, the sizes of the nanoparticles decreased to 209 nm, 204 nm, and 207 nm, respectively, and the size of Soot / PL35 was approximately 338 nm, which was found to be larger after coating (Fig. 1C). The dispersion values ​​of soot and Soot / Plu NP It was found that the size was manufactured uniformly with a size of less than 0.3 (Fig. 1D).

[0099] Dispersion stability: As can be seen from Fig. 2, in order to evaluate the dispersion stability of soot and Soot / Plu NP for 2 weeks, the yield of edible charcoal was analyzed using UV-Vis spectroscopy after 2 weeks of storage in DIW. While soot had the lowest yield due to low stability, Soot / PF127 NP had the highest absorbance, confirming the best dispersion stability.

[0100] Freeze-drying stability: As can be seen from Figure 3, in the case of soot, partial precipitation occurred after freeze-drying, making redispersion impossible, whereas in the case of Soot / PF68 NP, Soot / PF127 NP, and Soot / PP123 NP, the size and dispersion values ​​were confirmed to be stably maintained.

[0101] Storage stability: As can be seen from Fig. 4, when the size and dispersion values ​​of soot and Soot / Plu NPs stored in DIW and PBS were analyzed every 4 weeks, the size and dispersion values ​​of soot, Soot / PL35 NP, and Soot / PL81 NP increased, but the size and dispersion values ​​of Soot / PF68 NP, Soot / PF127 NP, and Soot / PP123 NP were all confirmed to be able to maintain a constant size and dispersion value for 4 weeks.

[0102]

[0103] 1-2. Manufacturing and Characterization of PF127-Coated Edible Charcoal Nanoparticles (Soot / PF127 NP) by Ratio

[0104] (1) Experimental method

[0105] Based on the characteristic evaluation according to Example 1, PF127, which was selected as the most optimal group among the five Pluronics, was used to manufacture Soot / PF127 NPs at various ratios. First, to compare the characteristics of Soot / PF127 NPs at various PF127 ratios, PF127 solutions of different concentrations (10, 25, 50, 100, and 500 mg) were prepared in 1 mL of DIW. Then, Soot / PF127 NPs were manufactured by reacting with 5 mg of edible charcoal using the same method as Example 1. At this time, the ratios of edible charcoal and PF127 were 1:2, 1:5, 1:10, 1:20, and 1:50. To evaluate the characteristics of the manufactured nanoparticles, photographs were taken and UV-vis spectra, size, and dispersion values ​​were analyzed. To evaluate the stability of Soot / PF127 NPs by ratio, the UV-Vis spectra of the nanoparticles were analyzed while stored in deionized water (DIW) for 2 weeks. In addition, the size and dispersion values ​​of the nanoparticles before and after lyophilization were analyzed to assess whether the nanoparticles could be easily stored in a powder state before use. The changes in the size and dispersion values ​​of Soot / PF127 NPs in DIW or PBS at 37°C for 4 weeks were analyzed to assess the in vivo stability of the nanoparticles.

[0106] 2-2. Results

[0107] Characterization: As can be seen in Fig. 5, all Soot / PF127 NPs were manufactured in a deep black color with edible charcoal well dispersed at all ratios (Fig. 5A). In particular, when the polymer ratio was 1:10, the yield of edible charcoal was good, and the intensity of the UV-Vis absorption peak was the highest (Fig. 5B). The size of the nanoparticles increased as the ratio of coated PF127 increased, and the size was manufactured between 206 nm and 220 nm (Fig. 5C). The dispersion values ​​were all less than 0.3, confirming that the sizes were manufactured uniformly (Fig. 5D). In the case of Soot / PF127 (1:50), the polymer concentration was too high to manufacture, making characterization impossible.

[0108] Dispersion stability: As can be seen from Fig. 6, when the UV-Vis spectra of Soot / PF127 NP at each ratio were analyzed after storage in DIW for two weeks, it was confirmed that Soot / PF127 NP (1:20) had the best dispersion stability.

[0109] Freeze-drying stability: As can be seen from Figure 7, when the freeze-drying stability of Soot / PF127 NPs by ratio was evaluated, it was confirmed that the size and dispersion values ​​of all groups were well maintained without change.

[0110] Storage stability: As can be seen from Figure 8, when the Soot / PF127 NPs were stored in DIW and PBS for 4 weeks at different ratios, the dispersion values ​​were all confirmed to be stable at less than 0.3 without any change in size.

[0111]

[0112] 1-3. In vitro cytotoxicity and biocompatibility evaluation of Soot / PF127 NP (1:20)

[0113] (1) Experimental method

[0114] To evaluate the cytotoxicity of Soot / PF127 NP (1:20), fibroblasts and NIH 3T3 cells were cultured in Dulbecco's Modified Eagle's Medium (DMEM) supplemented with 10% fetal bovine serum (FBS) and 1% penicillin-streptomycin (PS). The cultured cells were seeded into each well of a 96-well plate at 10,000 cells / well and cultured in an incubator at 37°C and 5% CO2 for 12 h. Then, the cells were treated with Soot / PF127 NP (1:20) at various concentrations (10, 200, 500 μg / mL) for 24 h, and the cell viability was analyzed using a CCK-8 kit. The control group (0 μg / mL) was treated with cell culture fluid. In addition, to observe cell morphology after treatment with Soot / PF127 NP (1:20), NIH 3T3 cells were cultured and treated with 0.1 mg / mL and 1 mg / mL of Soot / PF127 NP (1:20) for 24 hours. The negative control group was treated with cell culture medium, and the positive control group was treated with 10% DMSO.

[0115] (2) Results

[0116] As can be seen from Fig. 9, Soot / PF127 NP (1:20) had excellent biocompatibility and did not cause any cytotoxicity, and the cell viability was confirmed to be over 90% even at a high concentration of 500 μg / mL. In addition, as can be seen from Fig. 10, when compared to the control group after treatment with Soot / PF127 NP (1:20) with excellent biocompatibility, no change in cell morphology was observed, proving that it is a stable material for use as a semi-permanent cosmetic dye and further in the fields of medicine and cosmetics.

[0117]

[0118] 1-4. Evaluation of oxidative stress by in vitro induction of reactive oxygen radicals by Soot / PF127 NP (1:20)

[0119] (1) Experimental method

[0120] NIH 3T3 cells were cultured in DMEM supplemented with 10% FBS and 1% PS, and 10,000 cells were seeded into each well of a 96-well plate. After 12 hours of incubation, 200 μL of Soot / PF127 NP (1:20) was treated at various concentrations (100, 200, 500 μg / mL), and then cultured in an incubator for another day. The negative control group (0 μg / mL) was treated with cell culture medium, and the positive control group was treated with H2O2 (10 μM). The remaining samples were then washed with PBS to remove the remaining samples, and intracellular reactive oxygen radicals were analyzed using H2DCFDA (10 μM).

[0121] (2) Results

[0122] As can be seen from Fig. 11, when the amount of reactive oxygen radicals (ROS) existing in the existing cells was 100%, the ROS level of the positive control group treated with H2O2 was 124.4 ± 4.8%, indicating oxidative stress, whereas the ROS level did not increase when treated with Soot / PF127 NP (1:20).

[0123]

[0124] 1-5. In vivo dye properties of Soot / PF127 NP (1:20)

[0125] (1) Experimental method

[0126] To evaluate the in vivo dye properties of Soot / PF127 NP (1:20), 20 μL of Soot / PF127 NP (1:20) was administered intradermally to 8-week-old Sprague-Dawley rats. The original solution was administered to two sites on the left, and a 1 / 2 dilution was administered to two sites on the right. After 0, 4, and 8 weeks, photographs were taken to observe the dye properties, and H&E staining was performed to determine the presence of inflammation.

[0127] (2) Results

[0128] As can be seen from Fig. 12, both the original solution (left) and the 1 / 2 dilution (right) of Soot / PF127 NP (1:20) were found to maintain their color well for 8 weeks without fading or change after intradermal administration. In addition, as can be seen from Fig. 13, the H&E staining results confirmed that Soot / PF127 NP (1:20) was a safe material that did not cause any inflammatory reaction intradermally for 8 weeks. Therefore, the excellent tattoo retention ability and biocompatibility of Soot / PF127 NP (1:20) were proven.

[0129]

[0130] Example 2

[0131] 2-1. Manufacturing and Characterization of Polymer Coated Cosmetic Dye Materials

[0132] (1) Experimental method

[0133] To develop a semi-permanent cosmetic dye with excellent biocompatibility and stability, Pluronic F127 and polyvinylpyrrolidone (PVP; molecular weight = 10 kDa) were coated on BA (brown amber), GS (gold shrine), RA (red amber), and SS (silver shrine) as polymers, respectively. First, to compare the characteristics of nanoparticles for the two polymers, PF127 or PVP (10 kDa) was prepared at a concentration of 50 mg / mL in 1 mL of deionized water (DIW). The polymer solution was added to the cosmetic dye (5 mg), followed by 4 mL of DIW, and the mixture was stirred for 1 hour with rotatory shaking. Next, the reaction solution was transferred to a 20 mL vial and homogenized for 1 hour using an ultrasonic homogenizer (amplitude = 20%, 5 s on / 15 s off). The nanoparticles thus manufactured were freeze-dried for three days. To analyze the characteristics of the manufactured nanoparticles, their morphology was photographed, and their absorbance was measured using UV-Vis spectroscopy. The size and dispersion values ​​of the nanoparticles were analyzed using a Zetasizer (ELSZ-2000, Otsuka).

[0134] (2) Results

[0135] Absorbance: As can be seen from Fig. 14, four cosmetic dyes coated with PF127 or PVP (10 kDa) were well dispersed and manufactured after ultrasonic homogenization, and unique UV-Vis spectra were observed for each color of the nanoparticles. In particular, the absorbance of the cosmetic dye coated with PF127 was confirmed to be higher after purification, indicating that the cosmetic dye was better dispersed when coated with PF127 than when coated with PVP.

[0136] Size and dispersion values: As can be seen in Fig. 15, the size of the gold shrine coated with PF127 (GS / PF127) was 402 nm, and the size of the gold shrine coated with PVP (10 kDa) (GS / PVP10) was larger at 894 nm. The other three cosmetic dyes were found to be manufactured with similar sizes regardless of the polymer type. The sizes of BA / PF127 and BA / PVP10k were 481 nm and 459 nm, respectively, and the sizes of RA / PF127 and RA / PVP10k were 568 nm and 418 nm, respectively. In the case of silver shrine, the sizes of SS / PF127 and SS / PVP10k were manufactured with sizes of 368 nm and 304 nm, respectively, regardless of the polymer type (Fig. 15a). The dispersion values ​​of the nanoparticles were less than 0.4 for all but the unstable GS / PVP10k, which was manufactured with a large size, indicating that the nanoparticles were manufactured more uniformly (Fig. 15b). It was confirmed that coating the cosmetic dye with PF127 resulted in more stable and uniform nanoparticle formation compared to coating it with PVP.

[0137]

[0138] 2-2. Manufacturing and Characterization of PF127 Coating BA / PF127 by Ratio

[0139] (1) Experimental method

[0140] To compare the properties of BA / PF127 according to the PF127 coating ratio, PF127 solutions (25, 50, 100, 250 mg) were prepared in 1 mL of DIW. Then, BA / PF127 was prepared by reacting with 5 mg of brown amber using the same method as in Example 1. Bare BA was also prepared without polymer coating using the same method. At this time, it was confirmed that when the coating ratio was 1:50 or higher, it was difficult to handle and manufacture because it was higher than the solubility of PF127. To evaluate the properties of the manufactured nanoparticles, photographs were taken and UV-vis spectra, size, and dispersion values ​​were measured. In addition, to evaluate the stability of BA / PF127 according to the ratio, the changes in size and dispersion values ​​of BA / PF127 dispersed in DIW at 37℃ for 4 weeks were analyzed.

[0141] (2) Results

[0142] Absorbance, size, and dispersion values: As can be seen from Fig. 16, BA / PF127 was manufactured well-dispersed while maintaining its own color according to the coating ratio (Fig. 16a). In particular, it was confirmed that the lower the coating ratio, the better the brown amber was dispersed, resulting in a higher intensity of the UV-Vis absorption peak (Fig. 16b). In addition, the size of the nanoparticles increased as the ratio of the coated PF127 increased, and they were manufactured to be between approximately 400 nm and 520 nm (Fig. 16c). The dispersion values ​​were all less than 0.4, indicating that the sizes were manufactured uniformly (Fig. 16d).

[0143] Stability: As can be seen from Fig. 17, when BA / PF127 according to coating ratio was stored in DIW at 37℃ for 4 weeks, the dispersion values ​​were all stably maintained at less than 0.4 without any size change from 1:20 or higher, whereas Bare brown amber, BA / PF127 (1:5), and BA / PF127 (1:10) showed significant increases in size and dispersion values ​​over the 4 weeks of stability evaluation, indicating unstable existence. Therefore, it was confirmed that the dispersion stability and long-term stability were the best when the weight ratio of BA and PF127 was 1:20.

[0144]

[0145] 2-3. In vitro cytotoxicity and biocompatibility evaluation of BA / PF127 (1:20)

[0146] (1) Experimental method

[0147] To evaluate the cytotoxicity of BA / PF127 (1:20), fibroblasts and NIH 3T3 were cultured in DMEM (Dulbecco's Modified Eagle's Medium) supplemented with 10% FBS (fetal bovine serum) and 1% PS (penicillin-streptomycin). The cultured cells were seeded into each well of a 96-well plate at 10,000 cells / well and cultured in an incubator at 37°C and 5% CO2 for 12 hours. Then, BA / PF127 (1:20) was treated with various concentrations (0.1, 0.5, 1, 2, 4 mg / mL) for 24 hours, and cell viability was analyzed using a CCK-8 kit. At this time, the control group (CTL, 0 mg / mL) was treated with cell culture fluid. In addition, to observe cell morphology after BA / PF127 (1:20) treatment, L929 cells were cultured according to the biological evaluation criteria for medical devices (ISO 10993-5: 2009), and then treated with 0.1 mg / mL and 1 mg / mL of BA / PF127 (1:20) for 24 hours. At this time, the cell culture medium was treated as the negative control, and the eluate of a polyurethane film containing 0.1% zinc diethyldithiocarbamate was treated as the positive control.

[0148] (2) Results

[0149] As can be seen from Fig. 18, BA / PF127 (1:20) has excellent biocompatibility, does not cause any cytotoxicity, and the cell viability was over 90% even at a high concentration of 4 mg / mL. In addition, as can be seen from Fig. 19, unlike the positive control group in which cell death occurred, no change in cell morphology was observed after treatment with BA / PF127 (1:20), which has excellent biocompatibility, as in the negative control group. At this time, the cytotoxicity of BA / PF127 (1:20) was confirmed to be grade 0 according to the biological evaluation criteria for medical devices (ISO 10993-5: 2009), and therefore, it is expected to be safely utilized as a semi-permanent cosmetic dye in the fields of pharmaceuticals and cosmetics.

[0150]

[0151] 2-4. Evaluation of oxidative stress by in vitro induction of reactive oxygen radicals by BA / PF127 (1:20)

[0152] (1) Experimental method

[0153] NIH 3T3 cells were cultured in DMEM supplemented with 10% FBS and 1% PS, and 10,000 cells were seeded into each well of a 96-well plate. After 12 h of incubation, 200 μL of BA / PF127 (1:20) was treated at various concentrations (0.1, 0.5, 1 mg / mL) and incubated in the incubator for another day. The negative control group (0 μg / mL) was treated with cell culture medium, and the positive control group was treated with H2O2 (10 μM). After washing with PBS and removing the residual sample, intracellular reactive oxygen radicals were analyzed using H2DCFDA (10 μM).

[0154] (2) Results

[0155] As can be seen from Fig. 20, when the amount of reactive oxygen radicals (ROS) existing in the existing cells was 100%, the ROS level did not increase when treated with BA / PF127 (1:20), whereas the ROS level of the positive control group treated with H2O2 was 132.3 ± 10.7%, indicating that oxidative stress occurred. This demonstrated that BA / PF127 (1:20) is a cosmetic dye with high yield and stability and low cytotoxicity and oxidative stress.

[0156]

[0157] 2-5. In vivo dye properties of BA / PF127 (1:20)

[0158] (1) Experimental method

[0159] To evaluate the in vivo dye properties of BA / PF127 (1:20), 20 μL of BA / PF127 (1:20) was administered intradermally to 8-week-old Sprague-Dawley rats. Photographs were taken immediately after administration and 4 weeks after administration to determine the tattoo retention ability of the dye.

[0160] (2) Results

[0161] As shown in Figure 21, the color was stably maintained for four weeks after intradermal administration of BA / PF127 (1:20) without any color fading or change. Furthermore, it was confirmed to be a safe material that did not cause any inflammatory reaction within the skin.

[0162]

[0163] Research Project Information

[0164] [Ministry Name] Chungcheongbuk-do Provincial Office

[0165] [Research Project Name] Bioceramic Materials Company Technology Development Support Project

[0166] [Dedicated Organization] Korea Institute of Ceramic Engineering and Technology

[0167]

[0168] While specific aspects of the present invention have been described in detail above, it should be apparent to those skilled in the art that these specific descriptions are merely preferred implementation examples and are not intended to limit the scope of the present invention. Therefore, the substantial scope of the present invention is defined by the appended claims and their equivalents.

[0169] It is expected that the dye composition according to the present invention can be usefully used in cosmetic compositions, ink compositions (e.g., tattoo compositions), paint compositions, resin compositions, food compositions, etc.

Claims

1. Dye nanoparticles, and Comprising a pluronic polymer coated on the surface of the dye nanoparticles, A dye composition having improved stability and color retention.

2. In paragraph 1, A dye composition having improved stability and color retention, characterized in that the dye nanoparticles include edible charcoal (soot), brown amber (BA), gold shrine (GS), red amber (RA), silver shrine (SS), orange amber (OA), gold lustre (GL), light gold (LG), or a combination thereof.

3. In paragraph 2, A dye composition having improved stability and color retention, characterized in that the weight ratio of the edible charcoal nanoparticles and pluronic is 1:1 to 100.

4. In paragraph 2, A dye composition having improved stability and color retention, characterized in that the above Pluronic coated edible charcoal nanoparticles comprise edible charcoal and Pluronic PF127 in a weight ratio of 1:

20.

5. In paragraph 2, A dye composition having improved stability and color retention, characterized in that the weight ratio of the brown amber (BA), gold shrine (GS), red amber (RA), silver shrine (SS), orange amber (OA), gold lustre (GL) or light gold (LG) and the pluronic polymer is 1:1 to 40.

6. In paragraph 2, A dye composition having improved stability and color retention, characterized in that the dye composition comprises brown amber (BA) and pluronic polymer F127 in a weight ratio of 1:

20.

7. In paragraph 1, A dye composition having improved stability and color retention, characterized in that the dye composition is used for hair, skin, eyes, eyebrows or eyelashes.

8. In paragraph 1, A dye composition having improved stability and color retention, characterized in that the dye composition is used in a cosmetic composition, an ink composition, a paint composition, a resin composition or a food composition.

Images