A kind of preparation method of humanized active decellularized corneal stroma scaffold

A corneal stroma and decellularization technology, applied in the field of tissue engineering of biological corneal materials, can solve the problems of insufficient corneal transparency, lack of human corneal conformation, insufficient mechanical strength, etc. Wide source of raw materials and high transparency

Active Publication Date: 2019-10-25
陕西省眼科研究所
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0004] A new type of corneal material that uses biological materials as seed cells to be planted on xenogeneic corneal transplantation materials has attracted widespread attention. Commonly used biological materials include amnion, collagen, etc. CN201019018003 involves planting amnion epithelial cells and amnion stromal cells on animal corneas According to the method, the biocompatibility of the prepared tissue engineering corneal material is improved, but its mechanical strength is insufficient. At the same time, due to its lack of human corneal structure, the corneal transparency after transplantation is insufficient.

Method used

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  • A kind of preparation method of humanized active decellularized corneal stroma scaffold
  • A kind of preparation method of humanized active decellularized corneal stroma scaffold
  • A kind of preparation method of humanized active decellularized corneal stroma scaffold

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Embodiment 1

[0040] The preparation method of the humanized active decellularized corneal stromal scaffold of this embodiment comprises the following steps:

[0041] 1. Preparation of animal-derived decellularized corneal stroma carrier material:

[0042] Take the eyeballs of healthy pigs within 2 hours of execution, wash them alternately with normal saline and normal saline containing 300U / mL tobramycin, cut off the cornea, there is no sclera on the cut cornea, and then wash the cut cornea with antibiotic rinse with normal saline; cut the cornea after washing under aseptic conditions, the thickness of the cut cornea is 400 μm, and place the cut cornea in a sterile plate covered with saline gauze at 4°C;

[0043] Freeze and thaw the cornea in a sterile plate for 3 times. The specific process of the freeze-thaw treatment is: place the cornea in a -80°C refrigerator for 15 minutes, then take it out and let it slowly thaw at room temperature, and complete once when the cornea is completely th...

Embodiment 2

[0056] The preparation method of the humanized active decellularized corneal stromal scaffold of this embodiment comprises the following steps:

[0057] 1. Preparation of animal-derived decellularized corneal stroma carrier material:

[0058] Take healthy ostrich eyeballs within 2 hours of execution, wash them alternately with normal saline and normal saline containing 300 U / mL tobramycin, cut off the cornea, there is no sclera on the cut cornea, and then wash the cut cornea with antibiotic rinse with normal saline; cut the cornea after washing under sterile conditions, the thickness of the cut cornea is 200 μm, and place the cut cornea in a sterile plate covered with saline gauze at 4°C;

[0059] Freeze and thaw the cornea in a sterile plate once. The specific process of the freeze-thaw treatment is: place the cornea in a -80°C refrigerator for 20 minutes, then take it out and let it slowly thaw at room temperature, and complete once when the cornea is completely thawed. fre...

Embodiment 3

[0072] The preparation method of the humanized active decellularized corneal stromal scaffold of this embodiment comprises the following steps:

[0073] 1. Preparation of animal-derived decellularized corneal stroma carrier material:

[0074] Take the eyeballs of healthy pigs within 2 hours of execution, wash them alternately with normal saline and normal saline containing 300U / mL tobramycin, cut off the cornea, there is no sclera on the cut cornea, and then wash the cut cornea with antibiotic rinse with normal saline; cut the cornea after washing under aseptic conditions, the thickness of the cut cornea is 300 μm, and place the cut cornea in a sterile plate covered with saline gauze at 4°C;

[0075] Freeze and thaw the cornea in a sterile plate for 4 times. The specific process of the freeze-thaw treatment is: put the cornea in a -80°C refrigerator for 10 minutes, then take it out and let it slowly thaw at room temperature, and complete once when the cornea is completely thaw...

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Abstract

The invention discloses a preparation method of a humanized active decellularized corneal stroma scaffold, which comprises inoculating mesenchymal stem cells on an animal-derived decellularized corneal stroma carrier material for directional induction culture to obtain the humanized active decellularized corneal stroma carrier material. Cell corneal stroma scaffold; the specific operation steps include: step 1, rehydration of animal-derived decellularized corneal stroma carrier material; step 2, inoculating 1st to 4th generation mesenchymal stem cells isolated and cultured in vitro into step 1 for rehydration After the animal-derived decellularized corneal stromal carrier material, then add orientation induction medium, in CO 2 Oriented induction culture was carried out in an incubator to obtain humanized active decellularized corneal stromal scaffolds. The humanized active decellularized corneal stroma scaffold of the present invention has the characteristics of high transparency, good toughness, high tensile performance and good biocompatibility, and can promote the proliferation and differentiation of damaged corneal stem cells, thereby promoting the healing of damaged cornea.

Description

technical field [0001] The invention belongs to the field of tissue engineering of biological corneal materials, and in particular relates to a preparation method of a humanized active decellularized corneal stroma scaffold. Background technique [0002] The cornea is a transparent membrane located at the front of the eyeball, and is an important refractive medium of the optic system. Its function determines the clarity of vision. Corneal opacity caused by corneal damage is an irreversible change, and severe cases can lead to permanent visual impairment or even blindness. At present, there are about 60 million corneal blind patients in the world, of which there are about 4 million in my country. Corneal transplantation is an important means of recovery. However, due to the shortage of corneal donor materials, only about 5,000 corneal transplants can be performed in my country every year, and how to obtain corneal transplant materials has become an urgent problem to be solved...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): A61L27/38A61L27/36A61L27/50A61L27/54
CPCA61L27/3633A61L27/3641A61L27/3687A61L27/3691A61L27/3834A61L27/3839A61L27/50A61L27/54A61L2300/41A61L2430/16A61L2430/40
Inventor 刘先宁朱秀萍汪耀程燕潘士印肖湘华银勇杨华吴洁安娜王亚妮
Owner 陕西省眼科研究所
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