Preparation method and application of collagen-based composite corneal substitute capable of delivering endothelial inhibitory hormone

A substitute, collagen technology, applied in the field of preparation of semi-interpenetrating network gel, can solve the problems of corneal transplantation failure, affecting corneal transparency, etc., to promote corneal regeneration, easy processing and long-term storage and transportation, and improve stability. The effect of sex and mechanical strength

Inactive Publication Date: 2014-07-30
TIANJIN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Although the growth of corneal neovascularization is beneficial to the removal of pathogenic microorganisms and tissue repair, it seriously affects the transparency of the cornea, which is one of the reasons for the failure of corneal transplantation

Method used

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  • Preparation method and application of collagen-based composite corneal substitute capable of delivering endothelial inhibitory hormone
  • Preparation method and application of collagen-based composite corneal substitute capable of delivering endothelial inhibitory hormone
  • Preparation method and application of collagen-based composite corneal substitute capable of delivering endothelial inhibitory hormone

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preparation example Construction

[0025] Preparation of GMAA (reference Jianshu Li, Zhizhang Guo, Jianyu Xin, Guanglei Zhao, Huining Xiao. 21-Arm star polymers with different cationic groups based on cyclodextrin core for DNA delivery. Carbohydrate Polymers, 2010, 79, 277-283.):

[0026] 1. Dissolve 1g of arginine in 10ml of deionized water, adjust the pH to about 11 with 2mol / L NaOH solution, and inject nitrogen gas into the solution for 10min;

[0027] 2. Slowly add glycidyl methacrylate (GMA) to the aqueous solution of arginine under nitrogen protection environment, so that the molar ratio of GMA to arginine is 4:1, and continue to React overnight at room temperature;

[0028] 3. Use a separatory funnel to remove the unreacted GMA (upper layer) in the system, add ten times its volume of anhydrous ether to the solution and fully stir, use a separatory funnel to remove the ether phase (upper layer) after standing, and remove the lower layer of liquid After freeze-drying, methacrylic glycylated arginine monom...

Embodiment 1

[0031] The proportion of each component of the collagen corneal substitute is as follows:

[0032] Pig skin type Ⅰ collagen: GMAA (mass ratio) = 1:0

[0033] Coll-NH 2 :EDC:NHS (molar ratio)=1:1:1

[0034] The steps of preparing the composite corneal substitute with the above-mentioned components are as follows:

[0035] 1. Preparation of cross-linked collagen corneal substitute: Take 0.5g of 13.7% pig skin type Ⅰ collagen solution into the syringe, and connect another syringe to seal through the T-shaped container, push and pull repeatedly to mix evenly, and then use a micro-syringe to measure Inject crosslinking agents EDC and NHS, in which Coll-NH2:EDC:NHS=1:1:1 (molar ratio), push and pull repeatedly to mix evenly, and then add an appropriate amount of 2mol / L NaOH solution with a micro-syringe to adjust the pH to about 5.5. After mixing evenly, pour the solution into the mold, and then put it into the clamp to fix it, 100% humidity, react at room temperature for 16 hour...

Embodiment 2

[0038] The proportions of the components of the collagen / GMAA composite corneal substitute are as follows:

[0039] Pig skin type Ⅰ collagen: GMAA (mass ratio) = 1: 0.5

[0040] Coll-NH 2 :EDC:NHS (molar ratio)=1:1:1

[0041] Irgacure2959:GMAA (molar ratio)=0.02

[0042] The steps of preparing the composite corneal substitute with the above-mentioned components are as follows:

[0043] 1. Preparation of composite corneal substitute: Take 0.5g of 13.7% pigskin type Ⅰ collagen solution into the syringe, and connect another syringe to seal through a T-shaped container, push and pull repeatedly to mix evenly; the measured amount is pig skin type Ⅰ collagen : The GMAA monomer of GMAA=1:0.5 (mass ratio) is dissolved in water, and moved into the T-shaped container through the sealing gasket with a micro-syringe, repeatedly pushed and pulled to mix evenly, and then inject the cross-linking agent EDC, NHS and photoinitiator sequentially with a micro-syringe Irgacure2959, where Coll...

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Abstract

The invention discloses a preparation method and application of a collagen-based composite corneal substitute capable of delivering endothelial inhibitory hormone. The collagen-based composite corneal substitute is prepared by transfecting endostatin gene to mesenchymal stem cells (MSCs) on the surface of a collagen / glycidyl methacrylated arginine (GMAA) polymer semi-interpenetrating network corneal substitute. The collagen-based composite corneal substitute has high biocompatibility, can be used for inducing and promoting corneal regeneration, can be degraded along with the corneal regeneration, has high mechanical and optical properties, and has high biocompatibility with human eye tissues, and the introduction of a polymer network can be used for improving the stability and mechanical strength of the corneal substitute in collagenase. By inoculating bone marrow MSCs on the surface of the corneal substitute and transfecting the endostatin gene, endostatin proteins can be continuously detected in a culture medium in two weeks. The corneal substitute can be used for effectively combining gene transfection and tissue engineering and is used for ophthalmonogy; and the manufacturing method is simple, and the corneal substitute is easy to process, store for a long time and transport.

Description

technical field [0001] The invention belongs to the field of tissue engineering, and more specifically relates to a semi-interpenetrating network gel compounded with a collagen molecule capable of delivering endothelial inhibitory hormone and a glyceryl methacrylated arginine (Glycidyl Methacrylated Arginine, GMAA) polymer network. The preparation method of the gel, the multifunctional gel is used for the treatment of ophthalmology-related diseases through tissue engineering and gene transfection, and can be used as a potential corneal substitute. Background technique [0002] About 10 million people worldwide suffer from blindness caused by corneal damage. Corneal damage is generally caused by corneal diseases or external physical and chemical injuries, and has become the second leading cause of blindness worldwide after cataracts. At present, allogeneic keratoplasty provided by corneal donors is a relatively widely accepted treatment for irreversible corneal damage. Alth...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): A61K47/42A61L27/24A61L27/54A61K48/00C08F120/36C08F2/48C08J3/24A61P27/02
Inventor 刘文广刘源杨建海王宏博
Owner TIANJIN UNIV
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