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A deep skin regeneration agent prepared by mixing stem cell extracts and traditional Chinese medicine extracts and its preparation method and application

A technology of skin regeneration and extracts, applied in the field of biomedicine, to achieve the effect of inhibiting cell aggregation, promoting wound repair, and promoting the regeneration process

Active Publication Date: 2020-10-16
上海亚睿生物科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] However, there is currently no report on the treatment of diabetic foot with mesenchymal stem cell extract combined with traditional Chinese medicine extract. The new topical gel preparation of esmolol hydrochloride, the drug for ulcers, patent document CN101167875A, published on April 30, 2008, the disclosed traditional Chinese medicine preparation for treating diabetic foot, is still limited to the repair of diabetic foot skin damage

Method used

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  • A deep skin regeneration agent prepared by mixing stem cell extracts and traditional Chinese medicine extracts and its preparation method and application
  • A deep skin regeneration agent prepared by mixing stem cell extracts and traditional Chinese medicine extracts and its preparation method and application
  • A deep skin regeneration agent prepared by mixing stem cell extracts and traditional Chinese medicine extracts and its preparation method and application

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0066] Embodiment 1 Preparation of deep skin regeneration agent of the present invention (1)

[0067] Step 1: Purification of umbilical cord mesenchymal stem cells

[0068] 1) Take out the umbilical cord tissue of the newborn, remove blood vessels, epithelial membranes and blood clots, separate Wharton's blocks, and cut them into 1mm 3 After size, put it into a 10cm sterile petri dish; digest with DMEM medium containing 0.1% collagenase II; after filtration, use DMEM medium containing 10% fetal bovine serum FBS for subculture, and replace it with serum-free medium on the third day Subculture in DMEM medium, once every 2 days; take a bottle of cells from passage 3 for subsequent treatment;

[0069] 2) digest the cells with DMEM medium containing 0.05M EDTA, and wash with PBS solution;

[0070] 3) The cells were resuspended in serum-free DMEM medium, and lysed and crushed by ultrasonic (120W, 15S*20 times, 15S interval), the crushed product was centrifuged at 12000rpm for 15 m...

Embodiment 2

[0082] Embodiment 2 Cytotoxicity test of deep skin regenerating agent of the present invention

[0083] 1. Experimental method

[0084] 1) Primary culture of umbilical cord mesenchymal stem cells;

[0085] 2) The umbilical cord mesenchymal stem cells were mixed with 2*10 5 cells / ml into a 12-well plate for experiments, and adherent culture for at least 24 hours;

[0086] 3) Add the deep skin regeneration agent of the present invention (mass concentration is respectively 0%, 0.5%, 1%, 2%, 4%, 10% and 15%), and add the cell damage agent bleomycin as a control ;

[0087] 4) Perform flow cytometric detection on the cells to detect cell apoptosis and cell viability (CCK8 method).

[0088] 2. Experimental results

[0089] For the CCK8 experiment of mesenchymal stem cells, see the results figure 1 , it can be seen that the deep skin rejuvenation agent of the present invention has no cytotoxicity below 10%, while the deep skin rejuvenation agent of 4% has the best cell repair ef...

Embodiment 3

[0090] Example 3 The deep skin regeneration agent of the present invention inhibits the growth of fibroblasts and promotes the growth of neural stem cells and vascular endothelial cells in cell experiments

[0091] 1. Experimental method

[0092] 1) Primary culture of different cells, including fibroblasts, neural stem cells and vascular endothelial cells;

[0093] 2) Divide cells into 2*10 5 Spread into 12-well plates for experiments, 4 wells for each type of cells, and culture on the wall for at least 24 hours;

[0094] 3) Add the deep skin rejuvenating agent (mass concentration 4%) of the present invention to each kind of cells respectively, and add bleomycin (final concentration is 2 μ g / ml) as the cell injury agent, the specific adding method is:

[0095]

[0096] 4) Perform flow cytometry detection on all cells to detect cell apoptosis and cell viability (CCK8 method).

[0097] 2. Experimental results

[0098] The results of cell apoptosis in each group are shown ...

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Abstract

The invention relates to a deep skin regeneration agent prepared by mixing stem cell extract and traditional Chinese medicine extract as well as a preparation method and application thereof. The preparation method of the deep skin regeneration agent comprises the following steps: (1) culturing umbilical cord mesenchymal stem cells, and preparing the extracts by utilizing ultrafiltration and other technologies; (2) extracting general flavones in a traditional Chinese medicine composed of blue-lotus, roses, flos hibisci, aloe, astragalus and liquorice; and (3) mixing the stem cell extract and the traditional Chinese medicine extract. The deep skin regeneration agent can achieve obvious treatment effects on diabetic foot by promoting tissue regeneration and killing fibroblast, and is safe, high-efficiency, small in side effects, simple in production process and low in cost.

Description

technical field [0001] The invention belongs to the technical field of biomedicine, and specifically relates to a deep skin regeneration agent prepared by mixing stem cell extracts and traditional Chinese medicine extracts, a preparation method and application thereof. Background technique [0002] Diabetic foot, also known as diabetic foot ulcer, is a skin and neuropathy caused by diabetes. The main reason is the destruction of the soft tissue of the foot due to neuropathy, mainly manifested as mild sensory loss, ulcers and persistent infection, and in severe cases, it will cause joint disease, toe deformity and even amputation. At present, there is still no definite and effective method for the treatment of diabetic feet, because the sensory loss caused by peripheral nerve damage makes it difficult for patients to distinguish the damage to the feet, which in turn leads to infection and ulcers. Involuntary injuries and neuropathy can lead to vascular and muscle distortions...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): A61K36/896A61K36/886A61P17/02A61P3/10A61K35/28
CPCA61K9/0014A61K35/28A61K36/185A61K36/481A61K36/484A61K36/62A61K36/738A61K36/886A61K36/896A61K2300/00
Inventor 童强汪佳祺
Owner 上海亚睿生物科技有限公司
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