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Exogenous cornea substrate without cells and its preparation method and use

A corneal stroma, cell-free technology, applied in medical science, eye implants, prostheses, etc., can solve the problem of infectious pathogens infecting the human body

Inactive Publication Date: 2005-02-16
THE AFFILIATED SIR RUN RUN SHAW HOSPITAL OF SCHOOL OF MEDICINE ZHEJIANG UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, the strong rejection after xenotransplantation and the possibility of infectious pathogens infecting the human body after xenotransplantation have prevented xenotransplantation from being clinically applied so far. 8

Method used

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  • Exogenous cornea substrate without cells and its preparation method and use
  • Exogenous cornea substrate without cells and its preparation method and use
  • Exogenous cornea substrate without cells and its preparation method and use

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0023] Example 1 Preparation of decellularized porcine corneal stroma

[0024] Select healthy donor pigs in qualified farms, anesthetize with intramuscular injection of sodium thiopental, remove the pig’s eyeballs, trim the accessory tissues on the surface of the eyeballs, soak the eyeballs in 5% povidone-iodine for 3 minutes, and then use Rinse with 500 ml of normal saline, and store it in a freezer at -70°C until decellularization.

[0025] After the eyeball is thawed at normal temperature, cut off the whole cornea along the limbus under the operating microscope, scrape off the corneal epithelial layer and tear off the Descemet membrane, see Figure 1a , Figure 2a , Figure 3a , then place the corneal stroma in 30ml of ionic detergent, place it on a constant temperature shaker with 300 rpm uninterrupted vibration for 24 hours, change the solution once every 6 hours, and use 0.05% SDS (SIGMA chemical company, St.Louis, MO) was dissolved in deionized water, and 1mmol / L EDTA...

Embodiment 2

[0028] Example 2 Preparation of decellularized bovine corneal stroma

[0029] Choose healthy donor cattle in qualified farms, anesthetize with thiopental sodium needle intramuscularly, remove the eyeballs, trim the accessory tissues on the surface of the eyeballs, soak the eyeballs in 5% povidone-iodine for 3 minutes, and then use Rinse with 500 ml of normal saline and transport to the laboratory at 4°C.

[0030]Under the operating microscope, the whole cornea was cut along the corneal limbus, the epithelial layer was scraped off, and the corneal descemet membrane was torn off, and then the above-mentioned heterogeneous corneal stroma was placed in 30ml of non-ionic detergent, and 300 rpm was continuously shaken for 72 hours. The solution was changed every 12 hours. The non-ionic detergent was dissolved in deionized water with 0.5% Triton X-100 (SIGMA chemical company, St.Louis, MO), and 25mmol / L EDTA (SIGMA chemical company, St.Louis, MO) was added. , MO), the treatment temp...

Embodiment 3

[0033] Example 3 New Zealand Rabbit Therapeutic Corneal Transplantation Experiment

[0034] The acellular bovine corneal stroma prepared by the method of Example 2 was used as the donor material (hereinafter referred to as the donor), and healthy New Zealand rabbits were selected as the recipients for therapeutic penetrating keratoplasty, and the rabbits were successfully injected intramuscularly with thiopental sodium After general anesthesia, routine PVP iodine disinfection of the operated eye, laying a drape, opening the eyelids with a lid speculum, and fixing the upper and lower rectus muscle sutures, the donor cornea was rinsed with normal saline and placed on the corneal pad with the inner stroma facing upward, and a 7.5 mm ring was used to clean the eye. Drill in the center of the recipient cornea to the back of the anterior chamber, inject viscoelastic agent into the front, use corneal scissors to cut off the recipient cornea along the edge of the drill to prepare the i...

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Abstract

The invention provides a cell-less heterogenetic cornea material and manufacturing method and use, the manufacturing method for the product is: acquires the fresh animals' cornea at first; eliminates the cell membrane of the cornea material and nucleic acid with chemical method and enzyme method; carries on dehydration process to the cell-less heterogenetic cornea material; stores for backup. The method eliminates the antigenicity of the heterogenetic cornea material through eliminating the cell elements in the cornea material. At the same time, it reserves the integrity of glue fiber and transparency of the cornea. The material can be applied to construction of manmade cornea or used as the medical material for cornea transplantation, and cornea refraction operation directly.

Description

technical field [0001] The invention belongs to medical materials, and relates to a corneal stroma, in particular to a cell-free heterogeneous corneal stroma and a preparation method thereof. Corneal transplantation, refractive surgery. Background technique [0002] Corneal disease is the second leading cause of blindness worldwide. 1 For patients blinded by corneal disease, the only way to restore vision is through corneal transplantation. In my country, there are approximately 250,000 cases of blindness caused by corneal diseases every year, while only thousands of cases are received by corneal transplantation every year. The fundamental reason restricting the development of corneal transplantation is the extreme shortage of donor cornea sources. Therefore, it is the key to solve this contradiction between supply and demand to research and develop corneal substitutes that can replace human cornea. [0003] Artificial corneas have been studied for decades 2 , past rese...

Claims

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Application Information

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IPC IPC(8): A61F2/14A61L27/00A61L27/36A61L27/54
Inventor 姚玉峰朱也飞裘文亚
Owner THE AFFILIATED SIR RUN RUN SHAW HOSPITAL OF SCHOOL OF MEDICINE ZHEJIANG UNIV
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