Method for preparing carrier bracket of tissue engineering artificial corneal endothelium by using fresh amniotic membrane

Abstract

The invention relates to a method for preparing a carrier bracket of tissue engineering artificial corneal endothelium by using a fresh amniotic membrane. The method comprises the following steps: soaking and disinfecting the fresh amniotic membrane by tobramycin sulfate injection according to the mass ratio of 1:1000; after reverse digestion using trypsin-EDTA digestive juice, lightly scraping the epithelial surface of the amniotic membrane by a cell scraper to completely remove residual epithelial cells to obtain the denuded amniotic membrane which is tiled in culture plate holes for fixingand dry-posting; coating by special coating liquid for the denuded amniotic membrane; and sucking and then drying the coating liquid to obtain the carrier bracket of the tissue engineering artificialcorneal endothelium. The method has scientific and reasonable process, the prepared carrier bracket can be mass produced to meet the heavy demand of scale reconstruction of the tissue engineering artificial corneal endothelium and create conditions for sight rehabilitation of corneal endothelium blindness through clinical corneal transplantation, and the preparation method of the carrier bracket has low cost in in-vitro reconstruction and clinical treatment of the tissue engineering artificial corneal endothelium.

Description

technical field [0001] The invention relates to a method for preparing a tissue engineering artificial corneal endothelial carrier support by using fresh amniotic membrane, which belongs to the tissue engineering artificial corneal endothelial technology. Background technique [0002] The human corneal endothelium plays a vital role in maintaining corneal thickness, transparency and providing nutrients for the cornea. When the cornea is infected or traumatized by surgery, the corneal endothelial cells will be reduced to varying degrees. Once the density of corneal endothelial cells is lower than the critical density for forming a complete corneal endothelial layer and maintaining the physiological function of the corneal endothelium, the corneal endothelium will appear irreversible lesions. That is, corneal endothelial blindness. According to incomplete statistics, there are more than 800,000 patients with corneal endothelial blindness in my country at present, and most of ...

AI Technical Summary

Problems solved by technology

Scholars at home and abroad have used immortalized human corneal endothelial cells transfected with oncogenes, primary cultured human corneal endothelial cells, or human corneal endothelial cells cultured to the 4th or 5th passage in vitro as seed cells, respectively using the Descemet's membrane , de-epithelial amniotic membrane, collagen-chitosan blend membrane, polyhydroxyethyl acrylate, acellular porcine corneal stroma and N-(1-methylethyl)-2-acrylamide homopolymer as carriers Scaffolds successfully reconstructed human corneal endothelial tissue analogues in vitro in shape, transparency and tissue structure similar to normal corneal endothelium. Genetically immortalized cells are either primary cultured cells from eye bank corneas or only 4-5 passages in 24-well culture plates, tissue is limited due to potential tumorigenicity or extremely limited number of seed cells Scale in vitro reconstruction of engineered artificial corneal endothelium and its clinical application

Existing research reports on in vitro reconstruction of tissue-engineered artificial corneal endothelium are still limited to experimental research, and cannot meet the large number of clinical transplantation needs of more than 800,000 patients with corneal endothelial blindness in my country and more than 10 million patients worldwide.