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Biological amniotic membrane with complete cell structure and controllable degradation and preparation method thereof

An amniotic membrane and biological technology, applied in the field of biomedicine, can solve the problems of large cell proliferation and anti-inflammatory loss, easy destruction of amniotic membrane collagen and fibrous structure, reduced repair and anti-inflammatory effects, etc., and achieve good clinical treatment, cell and collagen structure. full effect

Pending Publication Date: 2021-07-13
北京桀亚生物医学研究有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Fresh amnion and refrigerated amnion can indeed retain a variety of collagen and active factor components to the greatest extent, but this type of amnion has potential safety hazards such as virus transmission and immunogenicity without virus inactivation, and fresh amnion may be attached to the outer layer of chorion. Or ingredients may hinder the effectiveness of the amnion
At present, the acellular amnion on the market usually uses one or more mixed solutions of acid, alkali, hypertonic saline solution, sodium lauryl sulfate or trypsin to remove cells and outer chorion. The method of soaking in chemical solvents is easy to damage the collagen and fiber structure of the amniotic membrane, and some factors that promote cell proliferation and anti-inflammation will lose a lot, making its repair and anti-inflammatory effects greatly reduced

Method used

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  • Biological amniotic membrane with complete cell structure and controllable degradation and preparation method thereof
  • Biological amniotic membrane with complete cell structure and controllable degradation and preparation method thereof
  • Biological amniotic membrane with complete cell structure and controllable degradation and preparation method thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0040] Embodiment 1 crosslinking agent is 0.05%, 0.1%, 0.25%, 0.5%, 1%, 2% glutaraldehyde solution

[0041] This embodiment provides the biological amniotic membrane obtained when the cross-linking agent concentration is 0.05%, 0.1%, 0.25%, 0.5%, 1%, and 2%; the specific steps are as follows:

[0042] (1) Spread the fresh amnion, and use a smooth sheet stainless steel scraper to manually scrape it against the growth direction of the chorion on the surface of the amnion to obtain amnion without chorion;

[0043] (2) Soak the amnion after scraping off the chorion in 3% hydrogen peroxide for 2 hours;

[0044] (3) Place the amniotic membrane soaked in hydrogen oxide in purified water for three consecutive washes, and then place it in 75% ethanol for 2 hours;

[0045] (4) Place the amniotic membrane treated with hydrogen peroxide and ethanol in purified water for three consecutive washes, and place them in cross-linking agent concentrations of 0.05%, 0.1%, 0.25%, 0.5%, 1%, and 2%,...

Embodiment 2

[0051] Embodiment 2 cross-linking agent is 0.1% glutaraldehyde solution

[0052] This embodiment provides the bio-amniotic membrane prepared when the cross-linking agent is 0.1% glutaraldehyde solution, and the specific steps are as follows:

[0053] (1) Spread the fresh amnion, and manually scrape it against the growth direction of the chorion on the surface of the amnion with a smooth sheet-shaped stainless steel scraper to obtain chorion-free amnion;

[0054] (2) Soak the amnion after scraping off the chorion in 3% hydrogen peroxide for 2 hours;

[0055] (3) Place the amniotic membrane soaked in hydrogen oxide in purified water for three consecutive washes, and then place it in 75% ethanol for 2 hours;

[0056] (4) Place the amniotic membrane treated with hydrogen peroxide and ethanol in purified water for three consecutive washes, then place it in a 0.1% glutaraldehyde solution, place it in a shaker, and set the shaker speed to 100r / min, the time is set to 1h, and the c...

Embodiment 3

[0059] Embodiment 3 cross-linking agent is 0.2% genipin solution

[0060] This embodiment adopts the same preparation method as in Example 2, the difference is that the cross-linking agent used is 0.2% genipin solution, the shaker speed is set to 50r / min, and the time is set to 30min for cross-linking treatment; Specific steps are as follows:

[0061] (1) Spread the fresh amnion, and use a smooth sheet stainless steel scraper to manually scrape it against the growth direction of the chorion on the surface of the amnion to obtain amnion without chorion;

[0062] (2) Soak the amnion after scraping off the chorion in 3% hydrogen peroxide for 2 hours;

[0063] (3) Place the amniotic membrane soaked in hydrogen oxide in purified water for three consecutive washes, and then place it in 75% ethanol for 2 hours;

[0064] (4) Place the amniotic membrane treated with hydrogen peroxide and ethanol in purified water for three consecutive washes, then place it in a genipin solution with ...

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Abstract

The invention relates to a biological amnion with a complete cell structure and controllable degradation and a preparation method thereof. The preparation method provided by the invention comprises the following steps: manually scraping the chorion of the amniotic membrane, and soaking the amniotic membrane without the chorion in hydrogen peroxide and ethanol. In order to control the degradation rate of the amnion, in the preparation method provided by the invention, glutaraldehyde, formaldehyde, genipin or a mixed solution of glutaraldehyde, formaldehyde and genipin with the concentration of 0.05-2.5% is selected to carry out crosslinking treatment on the amnion. By adopting the preparation method provided by the invention, acellular reagents such as acid, alkali, enzyme, hypertonic salt and the like are not used, and a complete cell structure is kept; and the degradation rate of the amnion is controlled by controlling the use concentration of the cross-linking agent. During clinical application, amniotic membrane products with different crosslinking degrees and degradation times are selected according to different diseases.

Description

technical field [0001] The invention relates to the technical field of biomedicine, in particular to a biological amniotic membrane with complete cell structure and controllable degradation and a preparation method thereof. Background technique [0002] The amniotic membrane is located on the surface of the fetal chorion. It is a smooth, non-vascular, non-nerve, and lymphatic transparent film with a thickness of about 0.02-0.50 mm. It is composed of amniotic epithelial cells, basement membrane and matrix. Amniotic membrane has low immunogenicity, has the functions of reducing inflammatory response and inhibiting fibrous tissue proliferation, and can be used as an ideal tissue engineering repair material. Amniotic membrane is an immune-privileged biological graft. Natural amniotic membrane has a large number of different protein components such as collagen, fibronectin, laminin and proteoglycan. In addition, natural amnion contains a variety of nutritional factors, such as t...

Claims

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Application Information

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IPC IPC(8): A61L27/36A61L27/22A61L27/50A61L27/54A61L27/58
CPCA61L27/3604A61L27/50A61L27/58A61L27/227A61L27/54A61L2430/34A61L2300/414
Inventor 孙继煌张嘉楠
Owner 北京桀亚生物医学研究有限公司
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