Cosmetic composition and pharmaceutical composition for alleviating atopic dermatitis, hair loss, and wounds or reducing skin wrinkles

A cosmetic composition, a technology for atopic dermatitis, applied in the field of cosmetic compositions and pharmaceutical compositions for relieving atopic dermatitis, hair loss and trauma or reducing skin wrinkles, can solve problems such as insufficient research, and achieve disease Excellent, symptom-improving, and expression-raising effects

Pending Publication Date: 2020-09-25
GREEN CROSS HEALTH SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, studies on secreted substances other than NK cells themselves (i.e. secretions), their new uses, etc. are still insufficient

Method used

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  • Cosmetic composition and pharmaceutical composition for alleviating atopic dermatitis, hair loss, and wounds or reducing skin wrinkles
  • Cosmetic composition and pharmaceutical composition for alleviating atopic dermatitis, hair loss, and wounds or reducing skin wrinkles
  • Cosmetic composition and pharmaceutical composition for alleviating atopic dermatitis, hair loss, and wounds or reducing skin wrinkles

Examples

Experimental program
Comparison scheme
Effect test

Embodiment

[0073] Example: Obtaining Natural Killer Cell Secretions

[0074] 1. Obtaining Natural Killer Cell Secretions

[0075] 1-1. Preparation of CD3(-) PBMC seed cells

[0076] PBS (Phosphate Buffered Saline, LONZA, 17-516Q) was added to PBMCs obtained from healthy donors at a ratio of 1:1, followed by centrifugation at 1500 rpm and 4°C for 10 minutes. Divide PBMC clumps in 10 x 10 6 Cells / mL were suspended in MACS running buffer (PBS containing 2% FBS and 2 mM EDTA) and cell counts were determined using an automated cell counter.

[0077] In order to obtain seed cells depleted of CD3(+) cells, 5-10×10 7 Each cell was transferred to a 50 mL tube and centrifuged at 1200 rpm and 4°C for 10 minutes. Add 400 to 800 μL of MACS running buffer and 100 to 200 μL of CD3 magnetic beads (Miltenyi Biotech, 130050101) to 5-10 × 10 7cells in PBMC cells, and then reacted at 4°C for 20 minutes. After washing with more than 10 times more MACS running buffer, the product was centrifuged at 1350...

experiment example 1

[0087] Experimental example 1. Evaluation of the cell proliferation ability of NK-CM

[0088] In order to evaluate the difference in proliferation ability of human fibroblasts between NK cell medium and NK-CM, Cell Counting Kit-8 (CCK-8) was used to determine cell proliferation ability.

[0089] Human fibroblasts (HDF-N) were suspended as single cells in 5 × 10 3 Cells / well were seeded in a 96-well plate and incubated at 37°C. After keeping the cells starved for 24 hours, the cells were treated with NK cell medium and NK-CM, respectively, and then further cultured for 24 and 48 hours. In order to determine the cell proliferation ability, the Cell Counting Kit-8 reaction was diluted 1 / 10 in supplement-free medium, then the diluted reaction was applied to the wells at 100 μl per well, and the reaction 1 Hour. The absorbance of the product at 450 nm was measured by an ELISA microplate reader (type 680, BIO-RAD). As a positive control, a medium supplemented with 10% FBS was us...

experiment example 2

[0091] Experimental example 2. Evaluation of the cell proliferation ability of NK-CM

[0092] Cell migration and wound recovery abilities of human fibroblasts in NK-CM were evaluated and determined using cell scratch assay.

[0093] Divide HDF-N cells at 1.0 × 10 4 Cells / insert were seeded in ibidi culture inserts at the bottom of 60mm culture dishes (μ-Dish 35mm, High Culture-Inserts; Thistle Scientific Ltd, UK) and cultured for 24 hours under cell culture conditions. After removing the chamber, the samples were diluted in the culture medium at the appropriate concentration, and the cells were treated with the diluted samples and incubated under cell culture conditions for 48 hours. After 24 and 48 hours, the migration ability of the cells was photographed by an electron microscope (Eclipse TS100, Nikon Instruments Inc., NY, USA).

[0094] Such as figure 2 As shown in (A), it can be seen that compared with the control, at 24 and 48 hours, as the concentration in the NK-CM...

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Abstract

The present invention relates to a cosmetic composition and a pharmaceutical composition which contain a secretion of natural killer cells so as to have excellent effects of increasing various factorsrelated to atopic dermatitis, hair loss, and wound alleviation or skin wrinkle reduction and improving the expression levels of the factors, thereby having excellent alleviation effects on the symptoms.

Description

technical field [0001] The present invention relates to a cosmetic composition and a pharmaceutical composition for improving atopic dermatitis, hair loss, wounds or skin wrinkles. Background technique [0002] Among the cells that form the immune system, natural killer cells (NK cells) in particular are cells that function at the forefront of the defense mechanism of the immune system in the body, thereby playing the role of removing tumor cells without prior sensitization by antigens , bacteria, intercellular parasites, or virus-infected host cells, reject inappropriate bone marrow transplantation, and modulate T-cell immune responses. [0003] Specifically, among such various functions of NK cells, since their ability to selectively kill cancer cells has been found, many related studies have been conducted. In the case of cancer patients, the number or activity of NK cells in peripheral blood is reduced compared with normal subjects. In animal experiments, it was found ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K8/98A61K35/17A61Q19/00A61Q19/08A61Q7/00
CPCA61K8/98A61K35/17A61Q7/00A61Q19/00A61Q19/08A61K8/64A61P17/02A61K38/2013A61K2300/00A61K8/981A61P17/14A61P17/00
Inventor 吴畅泽林玟姝金点溶朴宣奎李宗勋张珉廷金修爱林洙焕金凡峻朴东浩权泰隣罗正泰
Owner GREEN CROSS HEALTH SCI
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