Preparing method and application of acellular amniotic membrane used for repairing skin wound difficult to heal

A technology of wound repair and decellularization, applied in medical science, tissue regeneration, prosthesis, etc., can solve the problems of amnion matrix damage, basement membrane component damage, basement membrane damage, etc., to ensure integrity, promote regeneration and repair, Effect of improving decellularization efficiency

Active Publication Date: 2016-07-27
天晴干细胞股份有限公司
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, the existing amniotic membrane decellularization method still cannot guarantee the integrity of the amniotic matrix on the basis of complete removal of cells, especially it will cause serious damage to the basement membrane, affecting the adhesion and migration of seed cells or endogenous cells in vivo
Patent Publication No. CN1757717A (preparation method of dry active amniotic membrane) uses trypsin to treat for a long time in combination with cell scraper or cotton swab to remove epithelial cells, but long

Method used

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  • Preparing method and application of acellular amniotic membrane used for repairing skin wound difficult to heal
  • Preparing method and application of acellular amniotic membrane used for repairing skin wound difficult to heal
  • Preparing method and application of acellular amniotic membrane used for repairing skin wound difficult to heal

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specific Embodiment approach 1

[0033] Embodiment 1: A method for preparing acellular amniotic membrane for skin refractory wound repair according to this embodiment is characterized in that it is carried out according to the following steps:

[0034] 1. Select a full-term placenta, bluntly peel off the amniotic tissue, wash it with normal saline 2 to 3 times to remove impurities such as blood clots and colloids, and then soak it in normal saline containing antibiotics for 10 minutes for disinfection;

[0035] 2. Lay the amniotic membrane epithelium side up on the sterile nitrocellulose membrane, and let it dry for 1 to 2 hours under sterile conditions, so that the amniotic membrane and the nitrocellulose membrane are closely bonded to make an amniotic membrane patch and cut to an appropriate size. size;

[0036] 3. Put the amniotic membrane patch in the mixed digestion solution with a concentration of 0.25% to 0.5% trypsin and 0.2 to 0.5g / LEDTA·4Na in mass volume percentage, shake and digest at 37°C for 10 ...

specific Embodiment approach 2

[0044] Specific embodiment two: the difference between this embodiment and specific embodiment one is: the antibiotic content in the antibiotic saline described in step one is: concentration is the penicillin of 50mg / L, the streptomycin that concentration is 50mg / L and amphotericin B at a concentration of 2.5 mg / L. Others are the same as in the first embodiment.

specific Embodiment approach 3

[0045] Embodiment 3: This embodiment is different from Embodiment 1 in that: the airing time in step 2 is 1 to 1.5 hours. Others are the same as in the first embodiment.

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Abstract

The invention relates to a preparing method and application of an amniotic membrane, in particular to a preparing method and application of an acellular amniotic membrane used for repairing skin wound difficult to heal. The preparing method specifically comprises the steps of isolating, cleaning and sterilizing an amniotic membrane tissue; arranging and drying the amniotic membrane tissue on a nitrocellulose membrane to manufacture an amniotic membrane paster; vibrating and digesting the amniotic membrane paster in mixed digestive fluid of 0.25%-0.5% of pancreatin and 0.2-0.5g/L EDTA.4Na for 10-30 minutes at a temperature of 37 DEG C; rinsing the amniotic membrane paster in TE buffer solution; vibrating overnight to fully remove amniotic membrane cells in TE-TritonX-100 solution; washing the amniotic membrane paster for three times in TE solution; vibrating and degrading the amniotic membrane paster for 2-4 hours in nucleic acid degrading solution at a temperature of 37 DEG C; washing the amniotic membrane paster for three times in the TE solution. The prepared acellular amniotic membrane paster can be stored for a long time after being refrigerated, dried, packed and sterilized by cobalt 60 irradiation and is a tissue engineering material which can be taken and used when needed. The acellular amniotic membrane paster can be easily separated from the nitrocellulose membrane to be used for repairing the skin wound after rewatered.

Description

technical field [0001] The invention relates to a preparation method and application of amniotic membrane. Background technique [0002] Amnion tissue discarded during the perinatal period is one of the important sources of seed cells for regenerative medicine. It has a wide range of sources, is easy to obtain, and has no ethical and moral disputes. It has become a research hotspot in recent years. Amnion, as an immune-exempt biological graft, has biological properties such as anti-inflammation, anti-adhesion, pain relief, and promotion of epithelialization. It has been applied to burn patients and other surgical operations as early as 1910 to promote epidermalization, Reduce pain and prevent infection. In recent years, with the further understanding of amniotic membrane and the development of amniotic membrane preservation and preparation technology, amniotic membrane has once again become a research and application hotspot, and is widely used in corneal reconstruction, pl...

Claims

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Application Information

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IPC IPC(8): A61L27/36A61L27/60
CPCA61L27/3604A61L27/3687A61L27/3691A61L27/60A61L2430/40
Inventor 付寅生张怡孟庆雪刘艳青
Owner 天晴干细胞股份有限公司
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