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Method for preparing autologous beautifying micro-needle preparation and application thereof

A microneedle preparation and autologous technology, which is applied in the direction of medical formula, medical preparations containing active ingredients, drug delivery, etc., can solve the problems of insufficient wrinkle removal effect and poor stain lightening effect, so as to promote microcirculation, Effects of promoting wound healing and improving nutritional status

Pending Publication Date: 2017-05-24
SHENZHEN HORNETCORN BIOTECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0006] However, the existing cosmetic preparations such as platelet-rich plasma (PRP) are not effective in removing wrinkles, especially in reducing spots

Method used

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  • Method for preparing autologous beautifying micro-needle preparation and application thereof
  • Method for preparing autologous beautifying micro-needle preparation and application thereof
  • Method for preparing autologous beautifying micro-needle preparation and application thereof

Examples

Experimental program
Comparison scheme
Effect test

example 1

[0039] This example 1 is to isolate autologous PRP and peripheral blood mononuclear cells PBMCs:

[0040] Under sterile conditions, 20ml of peripheral venous blood was collected from the patient with a 20ml syringe, and anticoagulated with heparin sodium. Transfer the anticoagulated peripheral blood to a 50ml centrifuge tube at 400g and centrifuge for 15 minutes. After centrifugation, use a Pasteur pipette to collect the plasma layer and transfer it to a new 15ml centrifuge tube, and suck it up to 1-2 mm above the cell layer. 1500 g of autologous plasma containing platelets for 15 minutes. The upper layer of plasma was collected into a new 15ml centrifuge tube and separated from the lower layer of pellets (retain the bottom 3-5ml of plasma during separation and do not remove it from the tube). The remaining 3-5 ml of plasma and platelet pellet in the tube are resuspended to obtain concentrated PRP. Store the concentrated PRP and the remaining plasma at -80°C; dilute the lowe...

example 2

[0042] This example 2 is to induce fibroblasts, endothelial progenitor cells and immune cells in vitro:

[0043] The isolated PBMCs were resuspended in activated cell initial medium (containing activated cytokine solution), and the cell density was controlled at 1 × 10 6 cells / ml, transfer the cells into a T75 culture flask, shake well, and place them in a CO2 incubator with saturated humidity, 37°C, and 5% CO2 concentration.

[0044] After 2 days of culture, centrifuge at 500g / min for 6 minutes, collect the cells, resuspend the cell pellet with activated cell initial medium (containing activated cytokine solution), transfer it to a T75 culture flask, shake well, and place it in a CO2 incubator The culture was continued at medium saturated humidity, 37°C, and 5% CO2 concentration.

example 3

[0046] Example 3 is the quality inspection of the above-mentioned cultured cells

[0047]Morphology, purity, cell phenotype, cell number, viability, sterility, endotoxin, etc. were tested on the cells on the third day of culture. The specific operation includes the following steps: take 0.5ml of cell fluid to count with a cell counter, trypan blue staining to measure the cell viability, and flow cytometry to detect the cell phenotypes (CD45, CD3, CD56, CD34, CD31, CD42b, vimentin ( Vimentin)); the cell supernatant was collected by centrifugation, and the Mérieux automatic microbiological detection system was used to detect bacteria and fungi, LONZA mycoplasma detection kit to detect common mycoplasma, and Limulus reagent to detect endotoxin. After testing, on day 3, the number of cells was 2.72×10 7 cells (n=20), the cell viability by trypan blue staining reached 92.6% (n=20), and the cell phenotype detection of CD34+ cells reached 0.8%, of which endothelial progenitor cells ...

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Abstract

The invention belongs to the technical field of cosmetic mini-plastic surgery and specifically discloses a method for preparing autologous beautifying micro-needle preparation and application thereof. The preparation of the micro-needle preparation comprises the following steps: separating autologous platelet-rich plasma and peripheral blood mononuclear cells; culturing by using an autologous activated cell factor solution and a cell culture medium; replacing a fresh autologous activated cell factor solution and a cell culture medium the next day; collecting circulating fibrocytes, endothelial progenitor cells and immune cells on the fifth day, resuspending by using the autologous platelet-rich plasma and autologous plasma, thereby obtaining the autologous beautifying micro-needle preparation. Compared with the prior art, the method disclosed by the invention has the advantages that a composition of the circulating fibrocytes, endothelial progenitor cells, immune cells and platelet-rich plasma is used for cosmetic preparations, the circulating fibrocytes are capable of reconstructing skin tissues, secreting collagens and eliminating wrinkles; the endothelial progenitor cells can achieve the effects of local skin microcirculation and improving the skin nutrition state; the immune cells can achieve the effects of swallowing and taking away metabolic products; and the combination of the endothelial progenitor cells and the immune cells can achieve the effects of repairing spotted dull skin.

Description

technical field [0001] The invention belongs to the technical field of cosmetic micro-plastic surgery, in particular to a preparation method and application of an autologous cosmetic micro-needle preparation. [0002] technical background [0003] Facial anti-aging is an increasingly popular topic in cosmetic surgery. With the development of life science and the advancement of medicine, certain progress has been made in the prevention and anti-aging. The most direct manifestation of facial aging is the expression of wrinkles and pigmentation, and wrinkle removal and freckle removal have become a topic of increasing concern. [0004] With the improvement of people's living standards, people have higher requirements for spiritual living standards, requirements for temperament and appearance, and higher and higher requirements for beauty. Facial rejuvenation is to delay and resist the aging of facial skin through various methods, so that the face can maintain or even restore i...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K35/44A61K9/00A61P17/16A61P17/00A61K35/19A61K35/15A61K35/16
CPCA61K9/0021A61K35/15A61K35/16A61K35/19A61K35/44A61K2300/00
Inventor 王宇环付凡林科佳魏宗科罗晓玲
Owner SHENZHEN HORNETCORN BIOTECH
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