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In-vitro composition method for evaluating hair growing/hair loss prevention effect

A combination method, a hair loss prevention technology, applied in biochemical equipment and methods, microbial assay/inspection, compound screening, etc., and can solve problems such as limited predictive ability

Inactive Publication Date: 2018-09-14
黄健聪 +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

It has the advantages of high throughput, multiple endpoints, and high controllability of conditions, which is conducive to rapid and effective screening in the early stage of development, but a single method also has shortcomings such as limited predictive ability

Method used

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  • In-vitro composition method for evaluating hair growing/hair loss prevention effect
  • In-vitro composition method for evaluating hair growing/hair loss prevention effect
  • In-vitro composition method for evaluating hair growing/hair loss prevention effect

Examples

Experimental program
Comparison scheme
Effect test

example 1

[0103] Enzyme inhibition of example 1 ginger extract

[0104] In vitro model of S1,5α reductase inhibition

[0105] S1-1, reagent preparation: phosphate buffer (pH=7.0), enzyme extract, testosterone solution, NADPH solution, finasteride solution and Coomassie brilliant blue solution;

[0106] Preparation of Coomassie Brilliant Blue: Accurately weigh 100 mg of Coomassie Brilliant Blue G-250, add 50 mL of 95% ethanol, the solution turns blue; stir to dissolve, then add 100 mL of 85% (W / V) phosphate buffer solution with pH=7.0 and stir, the solution Blood red; finally distilled water to 1000mL, the solution turns brown; stir overnight on a magnetic heating stirrer, filter with filter paper for later use;

[0107] Preparation of protein standard solution: weigh bovine serum albumin to prepare a 0.1mg / mL protein standard solution, and store it in a refrigerator at 4°C;

[0108] The enzyme extract contains: 0.32mol / L sucrose, 1mmol / L 1,4-dithiothreitol, 0.1mmol / L EDTA, 10mmol / L Tr...

example 2

[0125] Example 2 Angelica extract cell efficacy analysis

[0126] S2, Cell efficacy analysis

[0127] S2-1, Cytotoxicity

[0128] S2-1-1, Reagent preparation: 10% FBS cell culture medium, 0.25% trypsin-0.02% EDTA, PBS and DMSO; MTT solution preparation: Weigh 0.5g of MTT and dissolve in 100mL of 0.01mol / L phosphate buffer Prepare a solution of 5 mg / mL, filter and sterilize with a 0.22 μm filter, aliquot, and store in the dark at 4°C;

[0129] S2-1-2 Angelica extract: take 100g of angelica medicinal material, wash it with boiling water 2-3 times, add ethanol of a certain concentration according to a certain ratio of solid to liquid, heat and reflux for extraction, filter, and collect the filtrate. Concentrate under reduced pressure to a certain crude drug concentration, add 1,3-butanediol and phenoxyethanol, mix well, centrifuge and pass through the membrane to obtain the Angelica sinensis extract. During the cytotoxicity test, 8 concentrations were set within the range of 0...

example 3

[0148] Example 3 Shouwu Extract Promotes Improvement of Blood Circulation

[0149] S3, Angiogenesis Analysis

[0150] S3-1, sample preparation

[0151] Shouwu extract: Take 100g of Shouwu medicinal material, wash it with boiling water for 2-3 times, add ethanol of a certain concentration according to a certain ratio of solid to liquid, heat and reflux for extraction, filter, and collect the filtrate. Concentrate under reduced pressure to a certain crude drug concentration, add 1,3-butanediol and phenoxyethanol, mix well, centrifuge and pass through the membrane to obtain the Shouwu extract.

[0152] S3-2, CAM preparation

[0153] SPF grade fertilized chicken embryos were incubated in an incubator (37.5°C, RH40-70%), turned over every day to check the development of the chicken embryos; on the seventh day, the well-developed chicken embryos were taken, and the eggshells were disinfected with alcohol and placed in the air chamber. Open an area of ​​about 2cm x 2cm; carefully ...

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Abstract

The invention discloses an in-vitro composition method for evaluating a hair growing / hair loss prevention effect. The in-vitro composition method comprises the following steps: S1, analyzing a 5alphareductase inhibition level; S2, carrying out cell effect analysis: analyzing cytotoxicity, a cell proliferation capability and an anti-oxidization capability; S3, analyzing angiogenesis; S4, evaluating a result and predicating. The in-vitro composition method disclosed by the invention is used for screening functionalized cosmetic raw materials through various biology methods and through a combined enzyme level test, a cell test and a tissue level test; a testing system is simple and low in price; special equipment is not needed; a detection is rapid, flexible and universal. The in-vitro composition method can be used for qualitatively and evaluating effects of hair growing raw materials; the method disclosed by the invention also can be used for replacing live animals and human volunteersand is used for screening and detecting the toxicity and effects of the hair growing and hair loss prevention raw materials.

Description

technical field [0001] The present invention relates to an in vitro combined method for evaluating hair growth / anti-hair loss efficacy, which can replace living animals for efficacy evaluation of new raw materials and new substances. Background technique [0002] Hair loss has become one of the common phenomena that plague modern people. According to relevant statistics, about 160 million people in my country are deeply troubled by hair loss. In terms of age, the youth born in the 90s and 80s have become the group most troubled by hair loss. It is reported that the global hair care industry is expected to develop at a compound annual growth rate of 4% between 2013 and 2018, and the market value will reach 60 billion US dollars by the end of 2018. [0003] The traditional hair growth evaluation method uses animal experiments, which have the disadvantages of large uncertainty, many influencing factors, long cycle, high cost, etc., and based on the requirements of animal welfa...

Claims

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Application Information

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IPC IPC(8): C12Q1/26C12Q1/02
CPCC12N2503/02C12Q1/26G01N33/5008G01N33/5014G01N33/5044G01N2333/90206G01N2500/10
Inventor 黄健聪程树军
Owner 黄健聪
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