Biomembrane for ocular surface restoration and preparation method thereof

A biofilm and ocular surface technology, applied in the field of tissue engineering medical biomaterials, can solve problems such as hidden dangers of virus transmission, difficult removal of amniotic membrane, loss of natural collagen, etc. effect of structure

Active Publication Date: 2014-10-08
SHAANXI BIO REGENERATIVE MEDICINE CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, since this kind of amniotic membrane has not been treated with virus inactivation, its sterile state cannot be ensured, and there is a risk of virus transmission.
In recent years, there are also some amniotic membrane products at home and abroad, all of which are freeze-dried, decellularized or air-dried at room temperature; During the drying and storage process, there are many factors that induce protein denaturation (such as the choice of cryoprotectant and lyoprotectant, the choice of freezing rate, etc.), which makes the freeze-drying process difficult to control, time-consuming and costly, and easily causes protein denaturation 1. The amniotic membrane becomes brittle, which causes the amniotic membrane to be easily torn during the operation, and needs to be replaced repeatedly, prolonging the operation time; and the method of air drying and air drying at room temperature makes the water volatilization process longer, and the effect of drying and dehydration is not ideal (generally, the moisture content >13 %),; In addition, the amniotic membrane loses a large amount of natural collagen after decellularization, especially the active factor components, which affects the clinical use effect
[0004] The amniotic membrane products disclosed in U.S. Patents US006152142A and US006326019B1 are blunt separation and removal of chorion after cesarean section placenta is cleaned, and the epithelium is spread upward on the nitrocellulose filter paper, and then pressed Preserved in DMEM:glycerol at a ratio of 1:1, stored in a -80°C refrigerator, the prepared amnion has not been decellularized, dried and sterilized, so the integrity of the amnion’s composition and structure is preserved to the greatest exten...

Method used

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  • Biomembrane for ocular surface restoration and preparation method thereof
  • Biomembrane for ocular surface restoration and preparation method thereof
  • Biomembrane for ocular surface restoration and preparation method thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0040] Step 1. Disinfection of raw materials: wash the amniotic membrane with physiological saline for 3 times, soak in 0.5M NaOH aqueous solution for 0.5 hour, and then wash with purified water for 3 times;

[0041] Step 2, antibacterial treatment: soak the sterilized amniotic membrane in physiological saline containing 50 μg / mL penicillin and 50 μg / mL streptomycin for 15 minutes;

[0042]Step 3, protection treatment: soak the amniotic membrane treated in step 2 in the mixed protection solution and shake for 24 hours; the mixed protection solution is a 1:1 mixture of dry protection solution and radiation protection solution by volume; the dry protection solution is 1g / L Trehalose aqueous solution of concentration; Radiation protection liquid is the ascorbic acid aqueous solution of 5g / L concentration;

[0043] Step 4. Vacuum drying: vacuum-dry the amniotic membrane treated in step 3, the vacuum degree is 0.1Mpa, and the temperature is controlled at 40°C, and the drying time i...

Embodiment 2

[0047] Step 1. Disinfection of raw materials: wash the amniotic membrane with normal saline for 5 times, soak in 75% alcohol solution for 5 hours, and then wash with purified water for 10 times;

[0048] Step 2, antibacterial treatment: soak the sterilized amniotic membrane in phosphate buffer solution with pH 7.0 containing 4 μg / mL gentamicin and 2.5 μg / mL amphotericin B for 1 hour;

[0049] Step 3, protection treatment: soak the amniotic membrane treated in step 2 in the mixed protection solution and shake for 1 hour; the mixed protection solution is a dry protection solution mixed with a radiation protection solution at a volume ratio of 1:2; wherein the dry protection solution has a volume concentration of 40% glycerol aqueous solution; the radioprotectant is 10g / L mannitol aqueous solution;

[0050] Step 4. Vacuum drying: the amniotic membrane treated in step 3 is vacuum-dried with a vacuum degree of -0.08Mpa, the temperature is controlled at 55°C, and the drying time is ...

Embodiment 3

[0054] Step 1. Disinfection of raw materials: wash the amniotic membrane 4 times with normal saline, soak in 1g / L hydrogen peroxide aqueous solution for 1 hour, and then wash 6 times with purified water;

[0055] Step 2, antibacterial treatment: soak the sterilized amniotic membrane in physiological saline containing 50 μg / mL penicillin and 2.5 μg / mL amphotericin B for 30 minutes;

[0056] Step 3, protection treatment: soak the amniotic membrane treated in step 2 in the mixed protection solution and shake for 24 hours; the mixed protection solution is a mixture of dry protection solution and radiation protection solution at a volume ratio of 1:1.5; the dry protection solution is 15g / L trehalose aqueous solution; radiation protection solution is 5g / L tea polyphenol aqueous solution.

[0057] Step 4. Vacuum drying: the amniotic membrane treated in step 3 is vacuum-dried with a vacuum degree of 0.05 Mpa, and the temperature is controlled at 45° C. for 2 hours.

[0058] Step 5, ...

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Abstract

The invention discloses a biomembrane for ocular surface restoration and a preparation method thereof. The biomembrane is a compact collagen membrane structure, comprises collagen I, collagen III and various active factors as main materials, has thickness of 0.01-0.05mm, tensile strength of 30-60MPa and stitching tearing force of 5-15N, and has effects of inhibiting scar and inflammation. Compared with the existing product, the biomembrane retains various natural components and a structure of amnion, has the same treatment effects and mechanical strength similar to that of fresh amnion and avoids virus hidden trouble of the biomembrane. The biomembrane is suitable for ocular surface burn and conjunctiva injury restoration, has good functions of promoting epithelium generation, resisting inflammation and inhibiting scar and has good treatment effects.

Description

technical field [0001] The invention belongs to the technical field of tissue engineering medical biomaterials, and in particular relates to a biofilm used for ocular surface repair and a preparation method thereof. Background technique [0002] Amnion, derived from cytotrophoblast, is the inner layer of the embryonic double membrane, which is composed of epithelial cell layer, basement membrane and avascular matrix. Amniotic membrane contains various collagens as well as laminin, fibronectin, glycosamine Polysaccharides, hyaluronic acid, etc. It is the presence of these active substances that can promote the adhesion and migration of epithelial cells, induce epithelial differentiation, and prevent epithelial apoptosis, making amnion a "transplantable basement membrane" to promote epithelialization. In addition, amnion stroma also contains many growth factors, such as nerve growth factor (NGF), hepatocyte growth factor (HGF), keratinocyte growth factor (KGF), transforming g...

Claims

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Application Information

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IPC IPC(8): A61L27/36A61L27/24A61L27/54
Inventor 刘博文刘影王二浩
Owner SHAANXI BIO REGENERATIVE MEDICINE CO LTD
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