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A kind of graft sheet for Descemet's membrane endothelial transplantation and its vacuole separation and preparation method

A technology of Descemet's elastic layer and graft sheet, which is used in eye implants, medical science, prosthesis, etc., can solve the problems of fragility and breakage, difficulty, thin thickness, etc., and achieves low price, large-scale industrialization, and a wide range of sources. Effect

Active Publication Date: 2018-05-04
GRAND LIFE SCI & TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The graft required by DMEK only includes Descemet's membrane and corneal endothelial layer, which is thin (about 15 μm), fragile, easy to break, and easy to curl, so it is difficult to completely separate it from the corneal stroma

Method used

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  • A kind of graft sheet for Descemet's membrane endothelial transplantation and its vacuole separation and preparation method
  • A kind of graft sheet for Descemet's membrane endothelial transplantation and its vacuole separation and preparation method
  • A kind of graft sheet for Descemet's membrane endothelial transplantation and its vacuole separation and preparation method

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0065] 1. Take the isolated porcine eyeball, and cut out the cornea with 1-2 mm scleral ring on the outer edge.

[0066] 2. Take the cornea obtained in step 1, and perform trypan blue staining on the scleral sulcus (in practice, the entire cornea obtained in step 1 can also be stained with trypan blue), then rinse with PBS buffer, and then use toothless Microscopic tweezers were used to scrape off the trabecular meshwork (pigment attached to the trabecular meshwork), exposing the end of the descemet membrane (Schwabe line).

[0067] The cornea after trypan blue staining at the scleral sulcus figure 1 .

[0068] Scraping the trabecular meshwork, exposing the cornea at the end of the descemet membrane figure 2 .

[0069] 3. Take the insulin syringe, draw out 0.6ml of corneal preservation solution, and then bend the needle into an angle of 90-120°.

[0070] For a photo of an insulin syringe with the needle bent at a 90-120° angle see image 3 .

[0071] 4. After step 3 is ...

Embodiment 2

[0093] Get the porcine DMEK graft piece that embodiment 1 prepares, detect cell density and live cell ratio.

[0094] The specific method is as follows:

[0095] 1. Take pig DMEK grafts and stain with 0.2% trypan blue staining solution for 2 minutes.

[0096] 2. After completing step 1, wash 2 times with PBS buffer.

[0097] 3. After completing step 2, stain with 1% Alizarin Red S staining solution for 3 minutes.

[0098] 4. After completing step 3, wash 2 times with PBS buffer.

[0099] 5. After step 4 is completed, observe under a microscope and count the cell density (3 high-power fields are randomly selected, and the sum is averaged) and the proportion of viable cells.

[0100] see photos Figure 11 . Arrows mark dead cells.

[0101] Get the porcine DMEK graft that embodiment 1 prepares, all reach following standard: cell density > 2500 / mm 2 , The proportion of viable cells >90%.

[0102] The graft prepared in Example 1 was used to perform Descemet's endothelial ke...

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Abstract

The invention discloses a graft for descemet membrane endothelial keratoplasty and a vacuole separation preparation method of the graft. The preparation method of the graft, provided by the invention, comprises the following steps: forming overall big vacuole between a stroma and a descemet membrane by a liquid injecting method, wherein porcine cornea serves as a raw material; then cutting and separating to prepare the graft consisting of the descemet membrane and an endothelium. The graft consisting of the descemet membrane and the endothelium is prepared from pig eyeballs, is expected to effectively relieve the current situation that human corneal endothelium donors are extremely insufficient and bring sight rehabilitating hope for a large number of patients suffering from corneal blindness, has important social value and very large industrialization and commercialization value, and will bring considerable economic value after being put into clinical market.

Description

technical field [0001] The invention relates to a graft sheet for descemet membrane endothelial corneal transplantation and a preparation method thereof for vacuole separation, in particular to a graft sheet for descemet membrane endothelial corneal transplantation prepared from porcine cornea and a preparation method for vacuole separation thereof. Background technique [0002] According to statistics, there are currently about 700,000 blind patients in my country due to severe corneal diseases, and corneal blindness has become the second treatable blindness. Corneal transplantation is the only means for patients to regain their sight. About 50% of patients can recover their eyesight through endothelial keratoplasty (EK). As the latest endothelial keratoplasty, Descemet Membrane Endothelial Keratoplasty (DMEK) has many advantages that other EKs do not have, so it has broad prospects for clinical application. However, the lack of human corneal donor sources seriously restr...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): A61F2/14
CPCA61F2/142A61F2240/001
Inventor 刘洋潘志强冯书堂
Owner GRAND LIFE SCI & TECH
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