A kind of preparation method of skin ulcer repair matrix

A skin ulcer and matrix technology, applied in medical science, prostheses, animal cells, etc., can solve problems such as high storage conditions and transportation requirements, limited wound repair effect, and inability to meet adjustments, so as to promote wound healing and reduce scar formation , the effect of prolonging the time of action

Active Publication Date: 2020-01-21
SHAANXI BOYU REGENERATIVE MEDICINE CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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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  • A kind of preparation method of skin ulcer repair matrix
  • A kind of preparation method of skin ulcer repair matrix
  • A kind of preparation method of skin ulcer repair matrix

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preparation example Construction

[0052] The preparation method of the skin ulcer repair matrix specifically adopts the decellularized, de-antigenized and thickened natural extracellular matrix as a biological scaffold, and the composite seed cells form a multi-layered cell membrane (having 2-10 cells) on the surface of the natural extracellular matrix. The skin ulcer repair matrix is ​​obtained after freeze-drying treatment; the seed cells involved are fibroblasts or embryonic or stem cells derived 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 and three-dimensional culture, freeze-drying, etc. The specific preparation process is as follows:

[0054] Step 1. Preparation of extracellular matrix:

[0055] Decellularization: The collagen membrane tissue used...

Embodiment 1

[0074] After the bovine pericardium was decellularized and washed, it was soaked in 1M sodium hydroxide solution for 5 min 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. The bovine pericardium was then immersed in PBS solution for 2 h. The bovine pericardium was cut into a circle of 10 cm, which was sterilized by irradiation with 10 kGy of cobalt 60 for use.

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

[0076] The sterilized bovine pericardium was spread on a sterile petri dish with a diameter...

Embodiment 2

[0079] After the bovine pericardium was decellularized and washed, 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. The bovine pericardium was then immersed in PBS solution for 6 h. The bovine pericardium was cut into a circle of 10 cm, which was sterilized by irradiation with 25 kGy of cobalt 60 for use.

[0080] The DMEM / F12 commercial medium and 10% fetal bovine serum were mixed as the basal solution according to the volume ratio of 9:1; then 60 ng insulin, 50 mg transferrin, 20 μg sodium selenite, and hydrocortisone were added according to the standard 500 ml basal solution. 100μg, epidermal growth factor 20μg, basic fibroblast growth factor 50ng, transforming growth factor beta 40ng, bovine pituitary extract 5mg, vitamin C 50mg.

[0081] The sterilized bovine pericardium was spre...

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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 of the skin loss damage, it needs to be treated in time. Small-area superficial skin damage can be repaired by the human body itself, but wounds larger than 20cm in diameter or deep to the dermis and chronic skin ulcers need to be treated by other means. [0003] 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 mellitus (DM) has been increasing year by year. At present, there are about 92 million diabetic patients in my ...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N5/071A61L27/36A61L27/38A61L27/60
Inventor 姚远
Owner SHAANXI BOYU REGENERATIVE MEDICINE CO LTD
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