A kind of decellularized biological amniotic membrane comprising loose layer and preparation method thereof

A technology of decellularization and loose layer, which is applied in the field of biological amnion and its preparation technology, can solve the problems of large loss of cell proliferation and anti-inflammation, reduced repair and anti-inflammatory effect, easy to be destroyed and removed, and achieve the improvement of repair and anti-inflammatory function , reduce loss, and ensure safety

Active Publication Date: 2021-05-11
CHENGDU QINGKE BIOTECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003] Fresh amniotic membrane and refrigerated amniotic membrane can indeed retain various collagen and active factor components to the greatest extent, but this type of amniotic membrane has not been treated with virus inactivation, decellularization, etc., and there are safety risks such as virus transmission and immunogenicity, and complete fresh amniotic membrane The ingredients contained are complex, and some structures or ingredients may hinder the efficacy of amnion, and the effect in clinical application may not be good; the existing amnion after virus inactivation, decellularization, descaling, etc. generally only contains basement membrane and dense layer, because the structure of the loose layer is loose, it is easy to be destroyed and removed during the treatment process, and some factors that promote cell proliferation and anti-inflammation lose a lot, making its repair and anti-inflammatory effects greatly reduced

Method used

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  • A kind of decellularized biological amniotic membrane comprising loose layer and preparation method thereof
  • A kind of decellularized biological amniotic membrane comprising loose layer and preparation method thereof
  • A kind of decellularized biological amniotic membrane comprising loose layer and preparation method thereof

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preparation example Construction

[0022] A method for preparing the decellularized biological amniotic membrane comprising a loose layer as described above, comprising the following steps:

[0023] Carry out virus inactivation to the washed biological amniotic membrane with peracetic acid-sodium chloride solution to obtain inactivated amniotic membrane;

[0024] Wash the inactivated amniotic membrane with normal saline and shake it, and wash it again with sodium bicarbonate solution to obtain the rewashed amniotic membrane;

[0025] The protease solution and the rewashed amnion are mixed and incubated, and the purified water is shaken and washed, and after being shaken and washed with a detergent, the decellularized biological amnion is prepared.

[0026] Further, in a preferred embodiment of the present invention, before virus inactivation, the freshly stripped amniotic membrane is washed with physiological saline, and then purified with peracetic acid-sodium chloride solution for 12-16 minutes at room temper...

Embodiment 1

[0040] This embodiment provides a method for preparing the decellularized bio-amnion that contains the loose layer. The method can prepare the decellularized bio-amnion that retains the loose layer, including the following steps:

[0041] 1.1 Wash the freshly stripped amniotic membrane twice with normal saline at room temperature;

[0042] 1.2 After cleaning, use 1.7mL / cm 2 Add a peracetic acid-sodium chloride solution with a peracetic acid content of 0.1% and a sodium chloride content of 0.8mol / L as a purification solution for virus inactivation, and purify by shaking at room temperature for 16 minutes;

[0043] 1.3 After the virus is inactivated, wash twice with normal saline at room temperature and rewash with 0.5% sodium bicarbonate solution to obtain rewashed amnion;

[0044] 1.4 Use 0.25% trypsin solution for 0.17mL / cm 2 The dosage is mixed with the rewashed amniotic membrane and incubated at 35°C for 150 minutes, and washed with pure water for 3 times;

[0045] 1.5 t...

Embodiment 2

[0047] This embodiment provides a method for preparing the decellularized bio-amnion comprising a loose layer, through which the acellular bio-amnion retaining the structure of the loose layer can be prepared, including the following steps:

[0048] 1.1 Wash the freshly stripped amniotic membrane with saline at room temperature for 3 times;

[0049] 1.2 After cleaning, use 1.7mL / cm 2 Add peracetic acid-sodium chloride solution with a peracetic acid content of 0.1% and a sodium chloride content of 1.2mol / L as a purification solution for virus inactivation, and shake and purify at room temperature for 12 minutes;

[0050] 1.3 After the virus is inactivated, wash with normal saline at room temperature for 3 times, and wash again with 10% sodium bicarbonate solution to obtain rewashed amniotic membrane;

[0051] 1.4 Use 0.4% trypsin solution for 0.24mL / cm 2 The amount of the amnion was mixed with the rewashed amnion and incubated at 39°C for 220 minutes, and washed with pure wat...

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Abstract

The invention provides a decellularized biological amniotic membrane containing a loose layer and a preparation method thereof, belonging to the field of biological amniotic membrane and its preparation technology. The preparation method of the decellularized biological amniotic membrane including the loose layer provided by the present invention has gone through steps such as decellularization and virus inactivation. On the premise of ensuring its safety, the natural structure of the amniotic membrane product is completely preserved, And the loose layer is also completely preserved, and the loose layer does not contain cells, the biological activity of its anti-inflammatory and repairing growth factors is retained to the greatest extent, and the loss of these growth factors is reduced, so that its repairing and anti-inflammatory functions are greatly improved. Improvement, so that it has better curative effect than ordinary acellular amniotic membrane products in the treatment of skin damage repair and anti-inflammation, reduces its immunogenic reaction, and has been inactivated by viruses, which improves safety and ensures the amniotic membrane It is clinically safe and effective.

Description

technical field [0001] The invention relates to the field of biological amniotic membrane and its preparation technology, in particular to a decellularized biological amniotic membrane comprising a loose layer and a preparation method thereof. Background technique [0002] Human amniotic membrane is derived from the innermost layer of the placenta, mainly composed of epithelial cell layer, basement membrane, compact layer, fibroblast layer and spongy layer. Type collagen and non-collagen components (laminin, nidogen, fibronectin), which are conducive to cell adhesion; the dense layer is one of the network fiber structures, containing type I collagen, type III collagen to form a bundle structure, and type V collagen, VI collagen Form a filamentous structure, the fascicle structure is conducive to maintaining the mechanical properties of the amniotic membrane, and the filamentous structure is connected with the basement membrane; the fibroblast layer is a loose collagen networ...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): A61K35/50A61P29/00A61P17/00
CPCA61K35/50A61P17/00A61P29/00A61L27/36A61L27/60
Inventor 康裕建丁雪琴张向飞付春燕王洁
Owner CHENGDU QINGKE BIOTECH
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