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Hydrogel-nanometer fiber membrane, preparation method and uses thereof

A nanofiber membrane and nanofiber technology, applied in the field of biomedicine, to achieve sufficient permeability, convenient surgical operation, and good permeability

Active Publication Date: 2013-04-03
EYECURE THERAPEUTICS INC JIANGSU
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] The purpose of the present invention is to provide a nanofiber polymer composite film and its preparation method and use for the deficiencies of the existing natural and synthetic carriers

Method used

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  • Hydrogel-nanometer fiber membrane, preparation method and uses thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0057] 1) Respectively mix 200μL chondroitin sulfate prepared above, 25μL H 2 O 2 Add 25μL horseradish peroxidase (HRP) to a 1.5mL centrifuge tube and mix well;

[0058] 2) Add the mixture prepared in step (1) to a 24-well cell culture plate, add 100μL to each well, at 37°C, 5% CO 2 Place in the incubator for 4 hours; add 0.5 mL of medium to each hole and put it in the incubator overnight;

[0059] 3) Aspirate the medium of the cell culture plate in step (2), spread 0.5mL Matrigel per well to facilitate the growth of RPE cells, put it in the incubator for 2 hours, and set aside;

[0060] 4) Aspirate the culture medium in the cultured RPE cell culture plate (calculated as 6-well cell culture plate), add 1mL PBS to each well to remove the remaining medium, after aspirating PBS, add 0.5mL per well Put 0.25% pancreatin-EDTA into the incubator for 2-5 minutes. After 85-95% of the RPE cells are detached from the plate wall, add 1 mL of medium to each well to stop the pancreatin-EDTA effect...

Embodiment 2

[0064] 1) Respectively mix 200μL chondroitin sulfate prepared above, 25μL H 2 O 2 Add 25μL horseradish peroxidase (HRP) to a 1.5mL centrifuge tube and mix well;

[0065] 2) Add the mixture prepared in step (1) to a 24-well cell culture plate, add 100μL to each well, at 37°C, 5% CO 2 Place it in the incubator for 3 hours; add 1 mL of medium to each hole and put it in the incubator overnight;

[0066] 3) Aspirate the culture medium of the cell culture plate in step (2), spread 1mL Matrigel per well to facilitate the growth of RPE cells, and put it in the incubator for 3 hours for use;

[0067] 4) Aspirate the culture medium in the RPE cell culture plate (calculated as the 6-well cell culture plate), add 0.5mL PBS to each well to remove the remaining medium, after aspirating the PBS, add 0.75mL per well It is 0.25% pancreatin-EDTA, put it in the incubator for 2-5 minutes, after 85-95% RPE cells are detached from the plate wall, add 1.5mL medium to each well to stop the pancreatin-EDTA e...

Embodiment 3

[0071] 1) Respectively mix 200μL chondroitin sulfate prepared above, 25μL H 2 O 2 Add 25μL horseradish peroxidase (HRP) to a 1.5mL centrifuge tube and mix well;

[0072] 2) Add the mixture prepared in step (1) to a 24-well cell culture plate, add 100μL to each well, at 37°C, 5% CO 2 Place it in the incubator for 5 hours; add 2 mL of medium to each hole and put it in the incubator overnight;

[0073] 3) Aspirate the culture medium of the cell culture plate in step (2), spread 2mL Matrigel per well to facilitate the growth of RPE cells, and put it in the incubator for 4 hours for use;

[0074] 4) Aspirate the culture medium in the RPE cell culture plate (calculated as the 6-well cell culture plate), add 2mL PBS to each well to remove the remaining medium, after aspirating the PBS, add 1mL per well to the content of 0.25 % Trypsin-EDTA, put it in the incubator for 2-5 minutes, after 85-95% of RPE cells are detached from the plate wall, add 2 mL of medium per well to stop the effect of t...

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Abstract

The present invention belongs to the field of biomedicine, and relates to a nanometer fiber polymer composite membrane, a preparation method and uses thereof. According to the nanometer fiber polymer composite membrane, the low layer is a hydrogel nanometer fiber membrane, the middle layer is a Matrigel matrix gel layer, and the upper layer is a retinal pigment epithelium layer. The nanometer fiber polymer composite membrane can be adopted as a bio-scaffold for supporting growth and transplantation of RPE cells, and has characteristics of softness, elasticity, good permeability, biodegradability and a structure similar to extracellular matrix, wherein adhesion and differentiation of the RPE monolayer cells and surgical procedure operations are easily achieved with the characteristics, and the nanometer fiber polymer composite membrane can be discharged out of a body through a metabolism process after transplantation so as not to generate harm to the human body. According to the present invention, a bio-nanometer material and cell engineering are combined, brightness and hope are provided for age-related macular degeneration (ARMD) patients with the successful research, and great economic benefits and social benefits are generated.

Description

Technical field [0001] The invention relates to the field of biomedicine, and relates to a nano-fiber polymer composite membrane and a preparation method and application thereof. Background technique [0002] Age-related Macular Degeneration (Age-related Macular Degeneration, AMD) is a multifactorial fundus disease, which is one of the common diseases that cause blindness in the elderly. This retinal degenerative disease is manifested as progressive loss of central vision, often leading to reading and other behavioral disorders. Although there is currently no specific treatment for this disease, scientists' research in the field of stem cells has brought the dawn of a cure for such patients, that is, transplanting retinal pigment epithelial cells (RPE) cultured on appropriate polymer scaffolds Into the eye to replace degenerated cells. This treatment program requires the stent to be biologically active and degradable, while the limited access to the subretinal space requires th...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61L27/44A61L27/38C12N5/071
Inventor 范国平薛志刚刘振山
Owner EYECURE THERAPEUTICS INC JIANGSU
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