Cross-linked decellularized amniotic membrane and its preparation method and application

A decellularization and amniotic membrane technology, which is applied in the field of cross-linked decellularized amniotic membrane and its preparation, can solve the problems of poor mechanical strength, biological stability and operability of decellularized amniotic membrane, so as to accelerate the proliferation rate in vitro and shorten the preparation time. , reduce the effect of wound shrinkage

Active Publication Date: 2019-06-21
GUANGZHOU RAINHOME PHARM&TECH CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the mechanical strength, biological stability and manipulability of acellular amnion are still poor

Method used

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  • Cross-linked decellularized amniotic membrane and its preparation method and application
  • Cross-linked decellularized amniotic membrane and its preparation method and application
  • Cross-linked decellularized amniotic membrane and its preparation method and application

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preparation example Construction

[0045] The cross-linked decellularized amniotic membrane of one embodiment, its preparation method comprises the following steps:

[0046] (1) Preparation of decellularized amniotic membrane:

[0047] The placentas of pregnant women who delivered by cesarean section were taken, and the infectious agent was ruled out by serum testing. The placentas used are selected from medically discarded placentas, which can be used for experimental research after being approved by the hospital and reviewed by the Medical Ethics Committee.

[0048] Under sterile conditions, peel off the amniotic membrane from the inner layer of the placenta, place it in sterile saline, rinse repeatedly to remove blood on the surface of the amniotic membrane, remove the chorion of the amniotic membrane, and rinse repeatedly with PBS solution with a pH of 7.2-7.4.

[0049] Soak the rinsed amniotic membrane in antibiotic solution for 3-7 minutes, and then soak in PBS with a pH of 7.2-7.4 for 8-12 minutes. This...

Embodiment 1

[0083] This embodiment provides a kind of decellularized amniotic membrane, and its preparation process is as follows:

[0084] (1) Take the placenta of the cesarean section pregnant woman, and exclude HIV, syphilis, HBV and HCV infection through serum testing.

[0085] (2) Peel off the amniotic membrane from the inner layer of the placenta under sterile conditions, place it in sterile normal saline, rinse repeatedly to remove the blood on the surface of the amniotic membrane, and bluntly remove the chorion of the amniotic membrane with forceps.

[0086] (3) After repeatedly rinsing the amniotic membrane with PBS solution with a pH of 7.3, soaking in antibiotic solution for 5 minutes and PBS solution for 10 minutes, this process was repeated 3 times.

[0087] (4) After re-rinsing with PBS solution, put the amniotic membrane into a solution containing DMEM and glycerol at a volume ratio of 1:1, and store at -80°C for 6 months to cover the window period of infection.

[0088] (...

Embodiment 2

[0091] This embodiment provides a cross-linked decellularized amniotic membrane, the preparation process of which is as follows:

[0092] Soak the decellularized amniotic membrane prepared in Example 1 in 30 mL of MES buffer for 1 h, then add EDC to make its concentration in MES buffer 0.05 mmol / mg, place it on a constant temperature shaker, set the speed at 30 rpm, Under the condition of 37°C, the cross-linking reaction was carried out for 5 minutes, washed with distilled water, and vacuum freeze-dried to obtain the product.

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Abstract

The invention relates to a crosslinked decellularized amnion, a composite skin substitute and a preparation method and application thereof. The preparation method for the crosslinked decellularized amnion includes the following steps: a decellularized amnion is immersed in an MES buffer solution, water-soluble carbodiimide, the concentration of which is 0.01mmol / mg to 0.1mmol / mg, is added, crosslinking reaction takes place for 1 to 10 minutes, and after washing and vacuum freeze drying, the crosslinked decellularized amnion is obtained. The crosslinked decellularized amnion prepared by the invention keeps the cytokines of the basilar membrane and moreover, the mechanical strength is enhanced. The composite skin substitute which is prepared from the decellularized amnion as a culture medium obtained by the invention can be used for wound repair.

Description

technical field [0001] The invention relates to the technical field of tissue engineering, in particular to a cross-linked decellularized amniotic membrane and its preparation method and application. Background technique [0002] Research on dermal substitutes has always been the focus and difficulty of tissue-engineered skin, and it is also the basis for the development of composite dermal substitutes. As a scaffold structure in the process of wound repair, dermal substitutes can not only promote the proliferation, migration and differentiation of epidermal cells, regulate the formation of basement membrane, but also guide the infiltration and proliferation of fibroblasts and vascular endothelial cells, deposit new collagen and form New blood vessels, thereby achieving the reconstruction of the dermal structure. [0003] Existing dermal substitutes can mimic the structural characteristics of normal dermis to a certain extent, but except for acellular dermis, they generally...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): A61L27/60A61L27/36C12N5/071
CPCA61L27/3604A61L27/3687A61L27/3691A61L27/60A61L2430/40C12N5/0625C12N2506/1392
Inventor 黄燕飞车七石刘少辉
Owner GUANGZHOU RAINHOME PHARM&TECH CO LTD
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