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Preparation method of skin ulcer repairing matrix

A skin ulcer and matrix technology, applied in medical science, prosthesis, etc., can solve the problems of high storage conditions and transportation requirements, limited wound repair effect, unsuitable for industrialization, etc., to promote wound healing, reduce scar formation, and apply wide range of effects

Active Publication Date: 2015-11-04
SHAANXI BOYU REGENERATIVE MEDICINE CO LTD
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AI Technical Summary

Problems solved by technology

[0009] The purpose of the present invention is to solve the problems of long cultivation period, high cost, complex process, unsuitable for industrialization, and inability to meet the role of adjusting the microenvironment of the wound surface in the existing tissue engineering skin construction method, so that its repair effect on the wound surface is limited, The disadvantages of short shelf life and high requirements for storage conditions and transportation

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  • Preparation method of skin ulcer repairing matrix
  • Preparation method of skin ulcer repairing matrix
  • Preparation method of skin ulcer repairing matrix

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preparation example Construction

[0052] The preparation method of the skin ulcer repair matrix specifically adopts the natural extracellular matrix after decellularization, antigen removal and thickening as a biological scaffold, and the compound seed cells form a multi-layered cell membrane (with 2 to 10 cells) on the surface of the natural extracellular matrix Cells with a layer of cell membrane), and the skin ulcer repair matrix can be obtained after freeze-drying; the seed cells involved are fibroblasts or embryos or stem cells from autologous or allogeneic sources.

[0053] According to the preparation process, the preparation method of the skin ulcer repair matrix mainly includes the steps of preparation of extracellular matrix, preparation of culture medium, separation and in vitro culture of seed cells, inoculation of cells, three-dimensional culture, and freeze-drying; the specific preparation process is as follows:

[0054] Step 1, preparation of extracellular matrix:

[0055] Decellularization: Aft...

Embodiment 1

[0074] After the bovine pericardium was decellularized and cleaned, it was soaked in 1M sodium hydroxide solution for 5 minutes at room temperature, and then washed with a large amount of 0.01M PBS solution until the pH was about 7. At this time, the thickness of bovine pericardium increased by 3 to 4 times. Then the bovine pericardium was submerged in PBS solution for 2h. The bovine pericardium was cut into a 10cm circle, irradiated with 10kGy cobalt 60 and sterilized for later use.

[0075] Mix DMEM / F12 commercial culture medium with 10% fetal bovine serum at a volume ratio of 9:1 as the base solution; then add 50 ng of insulin, 30 mg of transferrin, 10 μg of sodium selenite, and hydrocortisone according to the standard 500 ml base solution 60μg, epidermal growth factor 20μg, basic fibroblast growth factor 40ng, transforming growth factor β 20ng, adenine 10mg, vitamin C 30mg.

[0076] Spread the sterilized bovine pericardium on a sterile petri dish with a diameter of 10 cm...

Embodiment 2

[0079] After the bovine pericardium was decellularized and cleaned, it was soaked in 1‰ sodium hypochlorite solution at room temperature for 2 hours, and then washed with a large amount of 0.05M PBS solution until the pH was about 7. At this time, the thickness of bovine pericardium increased by 3 to 6 times. Then the bovine pericardium was submerged in PBS solution for 6h. The bovine pericardium was cut into a 10cm circle, sterilized by 25kGy cobalt 60 and then used for later use.

[0080] Mix DMEM / F12 commercial culture medium with 10% fetal bovine serum at a volume ratio of 9:1 as the base solution; then add 60 ng of insulin, 50 mg of transferrin, 20 μg of sodium selenite, and hydrocortisone according to the standard 500 ml base solution 100μg, epidermal growth factor 20μg, basic fibroblast growth factor 50ng, transforming growth factor β40ng, bovine pituitary gland extract 5mg, vitamin C 50mg.

[0081] Spread the sterilized bovine pericardium on a sterile petri dish with...

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Abstract

The invention provides a preparation method of a skin ulcer repairing matrix. According to the preparation method, a natural extracellular matrix subjected to decellularization, antigen removal and thickening treatment is adopted as a biological scaffold, seed cells are stratified on the surface of the natural extracellular matrix to form a cell membrane with multi-layer cells, and the skin ulcer repairing matrix is obtained after freeze drying treatment. By adopting the preparation method, the effects of surface cell stratifying of the extracellular matrix and partial cell ingrowth in the extracellular matrix are achieved; after the skin ulcer repairing matrix is used, the wound repairing can be effectively protected and promoted, and little scar is formed; the skin ulcer repairing matrix is short in preparation period, simple in process, suitable for industrialization, wide in application range, and good in wound repairing effect; the elasticity and toughness of the healed wound are close to those of the normal skin. By adopting freeze drying sterilization treatment, the product is guaranteed to have the expected use effect, the quality guarantee period is prolonged, and the transportation and storage requirements are reduced greatly.

Description

technical field [0001] The invention belongs to the technical field of tissue engineering and relates to a preparation method of a skin ulcer repair matrix. Background technique [0002] The skin is the first defense barrier of the human body and has an important function of maintaining the body's homeostasis. Therefore, no matter what the cause is, the skin loss damage needs to be treated in time. Small superficial skin injuries can be repaired by the body's own regulation, but patients with wounds larger than 20 cm in diameter or deep into the dermis and chronic skin ulcers need to be treated by other means. [0003] Chronic skin ulcer (chronic skin ulcer, CSU), also known as refractory ulcer, is a common clinical disease. Diabetic foot and venous ulcers are refractory ulcers. In recent years, with the improvement of people's living standards and the aging of society, the incidence of diabetes (diabctesmellitus, DM) has been increasing year by year. At present, there ar...

Claims

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Application Information

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IPC IPC(8): A61L27/36A61L27/38A61L27/60
Inventor 姚远
Owner SHAANXI BOYU REGENERATIVE MEDICINE CO LTD
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