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Biological amnion and preparation method thereof

A kind of amniotic membrane and biological technology, applied in the direction of medical science, surgery, layered products, etc., can solve the problems of unimproved single-layer amniotic membrane, the natural structure of amniotic membrane is greatly damaged, and the arrangement of collagen bundles is irregular, so as to increase product thickness and reliability. Operability, good decellularization effect, and simple preparation method

Active Publication Date: 2014-01-22
SHAANXI RUISHENG BIOTECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0004] Patents CN1369555A and CN102327640A respectively use reagents such as trypsin and TBS-SDS (TBS buffer containing sodium dodecyl sulfate) to decellularize the amnion. The amnion material prepared is relatively thin, easy to curl, and has poor adhesion. easy to slide off
In order to overcome the above disadvantages, the patent CN100368534C adopts cross-linking technology to prepare composite biologically derived amnion to improve its operability. After the step, the natural structure of the amniotic membrane is greatly damaged, resulting in irregular arrangement of collagen bundles and fracture of collagen fibers.
Aiming at the shortage of bio-derived amniotic membrane, patent CN1943796A discloses a method for making an irradiation cross-linked porous network decellularized amniotic membrane hydrogel auxiliary material, and proposes the concept of hydrogel dressing, but uses trypsin and ethylenediaminetetraacetic acid (EDTA) decellularizes the amniotic membrane, which can easily destroy the natural structure of the amniotic membrane, cause collagen fiber breakage, and reduce the mechanical properties of the material. The safety of the product is often accompanied by some side reactions during the combination process, and the two-time irradiation process will greatly destroy the spatial collagen structure of the natural amniotic membrane
Patent CN101040616A soaks amniotic membrane and growth factor to prepare drug amniotic membrane, but it still does not improve the operational shortcomings of single-layer amniotic membrane being thin and easy to curl, and the growth factor after soaking is only attached to the surface of the amniotic membrane, and the slow-release performance is poor

Method used

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  • Biological amnion and preparation method thereof
  • Biological amnion and preparation method thereof
  • Biological amnion and preparation method thereof

Examples

Experimental program
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Effect test

Embodiment 1

[0033] Step 1. Raw material pretreatment: remove surface blood stains from fresh amnion, and wash with phosphate buffer;

[0034] Step 2. Virus inactivation: soak the amniotic membrane in step 1 in 75% ethanol solution at room temperature 25°C for 2 hours, and wash with purified water 4 times;

[0035] Step 3, decellularization treatment: place the amniotic membrane in step 2 in a hypertonic solution (the hypertonic solution is a mixed solution of 4M NaCl and 0.05M NaOH) at a room temperature of 25°C, shake at room temperature for 2.5 hours, and then quickly wash with purified water. Then use purified water to shake at room temperature for 2.5 hours; repeat once to prepare the amnion layer;

[0036] Step 4. Collagen layer preparation: wash the amnion layer prepared in step 3 and place the epithelial surface (smooth side) down on the bottom of a container with the same size as the amnion, add collagen solution, the thickness is about 10.0 μm, -20 Pre-freeze at ℃ for 2 hours an...

Embodiment 2

[0041] Step 1. Raw material pretreatment: remove surface blood stains from fresh amniotic membrane and wash with purified water;

[0042] Step 2. Virus inactivation: soak the amniotic membrane in step 1 in 0.5% peracetic acid solution for 10 minutes at room temperature 25°C, and wash with purified water 4 times;

[0043] Step 3, decellularization treatment: place the amniotic membrane in step 2 at room temperature 25°C in a hypertonic solution (the hypertonic solution is a mixed solution of 1M NaCl and 0.10M NaOH), shake at room temperature for 0.5h, wash it with purified water quickly, and then Use purified water to shake at room temperature for 0.5 h; repeat 5 times to prepare the amnion layer;

[0044] Step 4. Preparation of collagen layer: Wash the amnion layer prepared in step 3 and place the epithelial surface (smooth side) downward on the bottom of a container with the same size as the amnion, add collagen solution, the thickness is about 40.0 μm, -20 After pre-freezin...

Embodiment 3

[0049] Step 1. Raw material pretreatment: remove surface blood stains from fresh amniotic membrane and wash with normal saline;

[0050] Step 2. Virus inactivation: soak the amniotic membrane in step 1 in 75% ethanol solution for 3 hours at room temperature of 25°C, and wash with purified water for 5 times;

[0051] Step 3, decellularization treatment: place the amniotic membrane in step 2 in a hypertonic solution (hypertonic solution is a mixed solution of 3M NaCl and 0.01M NaOH) at room temperature at 25°C, shake at room temperature for 1 hour, wash with purified water quickly, and then use Shake the purified water at room temperature for 1 hour; repeat 4 times to prepare the amnion layer;

[0052] Step 4. Preparation of collagen layer: Wash the amnion layer prepared in step 3 and place the epithelial surface (smooth side) downward on the bottom of a container with the same size as the amnion, add collagen solution, the thickness is about 30.0 μm, -20 After pre-freezing at ...

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Abstract

The invention relates to a biological amnion and a preparation method thereof. The biological amnion has three layers including a slow release layer, an amnion layer and a collagen layer from top to bottom, wherein the slow release layer consists of collagen and biological active factors, the amnion layer consists of an amnion subjected to decellularization treatment, and the collagen layer is formed by freeze-drying and compounding collagen. The biological amnion is prepared through the steps of raw material pretreatment, virus inactivation, decellularization treatm. The biological amnion prepared by the invention has the characteristics of an effect of slowly releasing the active factors, low antigenicity on removing epithelial cells, convenience in product operation, difficulty in curling, good adhesiveness with surrounding tissues, difficulty in sliding and the like. Meanwhile, by using a process for performing the decellularization treatment on the amnion, disclosed by the invention, a natural compact collagen structure in the amnion can be effectively retained, the biological amnion can fully achieve a physical barrier effect after being applied to tenorrhaphy, and an animal experiment proves that the biological amnion can effectively achieve an effect of preventing tissue adhesion.

Description

technical field [0001] The invention belongs to medical biomaterial technology, in particular to a bio-amnion and a preparation method thereof. Background technique [0002] Tissue adhesions are abnormal structures formed by the combination of connective tissue fibrous bands and adjacent tissues or organs, which can cause various postoperative complications: for example, adhesions after tendon repair, seriously affecting the recovery of the function of the surgical site, and even Can cause loss of local function. [0003] Amniotic membrane is the inner layer of human fetal membranes, including the thickest basement membrane and avascular interstitium in the human body. The amniotic membrane has low immunogenicity and can reduce inflammatory reactions and inhibit fibrous tissue proliferation. It can be used as an ideal Tissue engineering repair materials. [0004] Patents CN1369555A and CN102327640A respectively use reagents such as trypsin and TBS-SDS (TBS buffer containin...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61L31/12A61L31/00A61L31/16A61L31/04B32B9/02
Inventor 李晶
Owner SHAANXI RUISHENG BIOTECH
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