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Preparation method of decellularized biological amnion

A technology of decellularization and amnion, applied in tissue regeneration, medical science, prosthesis, etc., can solve the problems of limited clinical application range, residual amnion cells, insufficient mechanical properties, etc., to eliminate or weaken antigenicity, retain biological activity, Effects in simple steps

Inactive Publication Date: 2017-05-31
WUHAN YIJIABAO BIOMATERIAL CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] However, the current treatment of amniotic membrane mostly involves the use of mechanical scraping methods, resulting in defects such as cell residues, insufficient mechanical properties, and short degradation cycle of the amniotic membrane, which limits its clinical application.
According to clinical reports, due to the existence of amnion cells, in ophthalmology, amniotic membrane grafts may have complications such as graft rejection, graft loss, displacement dissolution, infection, etc., and its safety has certain risks

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0026] Wash the fresh amniotic membrane with a large amount of normal saline until there is no residual blood, then soak it in a hydrogen peroxide solution with a mass-volume ratio of 5:1 and a mass fraction of 5% for 2 hours; then put the amniotic membrane into a mass-volume ratio of In 5:1 analytically pure acetone, shake at constant temperature in a water bath for 2 hours to remove fat components; then soak the obtained amniotic membrane in sodium lauryl sulfate solution with a mass volume ratio of 5:1 and a mass fraction of 0.15% 18 hours; then soak in alcohol with a mass volume ratio of 5:1 and a mass fraction of 75% for 18 hours to remove water, then rinse the amniotic membrane repeatedly with a large amount of purified water until the pH value of the final cleaning solution is between 6.5-7.5 .

[0027] The amnion after cleaning was spread flat on the stainless steel freeze-drying tray, frozen in an environment below -70°C for 24 hours, and then put into a freeze dryer ...

Embodiment 2

[0029] Wash the fresh amniotic membrane with a large amount of normal saline until there is no residual blood, and soak it in a hydrogen peroxide solution with a mass volume ratio of 6:1 and a mass fraction of 5% for 2 hours; then put the amniotic membrane into a mass volume ratio of In 6:1 analytically pure acetone, the water bath was shaken at constant temperature for 2 hours to remove the fat component; then the obtained amnion was soaked in sodium lauryl sulfate solution with a mass volume ratio of 6:1 and a mass fraction of 0.2%. 18 hours; then soak in alcohol with a mass volume ratio of 5:1 and a mass fraction of 75% for 18 hours to remove water, then rinse the amniotic membrane repeatedly with a large amount of purified water until the pH value of the final cleaning solution is between 6.5-7.5 .

[0030] The amnion after cleaning was spread flat on the stainless steel freeze-drying tray, frozen in an environment below -70°C for 24 hours, and then put into a freeze dryer...

Embodiment 3

[0032] Wash the fresh amniotic membrane with a large amount of normal saline until there is no residual blood, then soak it in a hydrogen peroxide solution with a mass-volume ratio of 6:1 and a mass fraction of 10% for 2 hours; then put the amniotic membrane in a mass-volume ratio of In 6:1 analytically pure acetone, shake at constant temperature in a water bath for 2 hours to remove fat components; then soak the obtained amniotic membrane in sodium lauryl sulfate solution with a mass volume ratio of 6:1 and a mass fraction of 0.25% 18 hours; then soak in alcohol with a mass volume ratio of 5:1 and a mass fraction of 75% for 18 hours to remove water, then rinse the amniotic membrane repeatedly with a large amount of purified water until the pH value of the final cleaning solution is between 6.5-7.5 .

[0033] The amnion after cleaning was spread flat on the stainless steel freeze-drying tray, frozen in an environment below -70°C for 24 hours, and then put into a freeze dryer f...

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Abstract

The invention provides a preparation method of a decellularized biological amnion. The preparation method comprises cleaning, chemical treatment and freeze drying of the amnion. According to the method, a fresh amnion is washed clean with normal saline, residual blood on the amnion is washed away, the obtained amnion is treated with hydrogen peroxide, acetone, sodium dodecyl sulfate and alcohol again, after alcohol treatment, the obtained amnion is subjected to profound hypothermia freezing in an environment at subzero 70 DEG C for 16-25 h, and the obtained amnion is placed in a freezer dryer for freeze drying according to a set freeze drying curve. A finished decellularized biological amnion is obtained after irradiation sterilization. The amnion is jointly treated with hydrogen peroxide, acetone, sodium dodecyl sulfate and alcohol, immunogenic ingredients of an epithelial cell layer, a fibrous membrane cell layer and a sponge layer in the amnion can be effectively removed, meanwhile, a basilar membrane and a compact layer of the amnion are reserved, and the biological activity and biomechanical properties of the amnion are better reserved; on the other hand, the immunogenicity of the decellularized biological amnion can be further reduced through freeze drying treatment, and the immunological rejection of host cells is reduced. In addition, the preparation method of the decellularized biological amnion is simple and easy to implement, and large-scale production operation is facilitated.

Description

technical field [0001] The invention relates to the field of biomaterials, specifically designing a method for preparing allogeneic implantable biomaterials. Background technique [0002] The amniotic membrane is the innermost layer of the fetal membrane, which belongs to a kind of biological membrane and is a kind of biologically derived material of natural extracellular matrix. The amniotic membrane is differentiated from the trophoblast layer, the surface is smooth, translucent, tough and elastic, and the thickness is 0.02-0.50mm. It is mainly composed of type I and type III fibrous collagen and contains a small amount of type VI and type V collagen. Growth factors, such as fibroblast growth factor-2 (fibroblast growth factor-2, FGF-2), transforming growth factor-β (transforming growth-β, TGF-β), angiogenesis-promoting vascular endothelial growth factor (VEGF ), also contains fibronectin (FN) and so on. Amniotic membrane does not express HLA-A, B, C and DR antigens or β...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61L27/36A61L27/54
CPCA61L27/3604A61L27/3683A61L27/3687A61L27/3691A61L27/54A61L2430/40
Inventor 万飞许世生唐小雄
Owner WUHAN YIJIABAO BIOMATERIAL CO LTD
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