Method for preparing decellularized cornea

Abstract

The invention provides a method for preparing decellularized cornea. Due to the selection of components and proportions, colloid osmotic pressure of a decellularized cornea preparation liquid can be maintained by using a prepared protection liquid, and compared with a conventional decellularized cornea preparation liquid, the decellularized cornea preparation liquid is capable of remarkably alleviating the situation that the cornea transparency is degraded as an extracellular matrix such as collagen is excessively swelled and lost in the later cell treatment process; in addition, due to the addition of antibiotics such as tobramycin, bacterium propagation of susceptible cornea can be inhibited, and the contamination rate in the treatment process can be reduced. By adopting the method provided by the invention, cells can be crushed, and the situation that the trenchancy is degraded as cornea matrix collagen is damaged by long-term high pressure can be avoided. Secondly, as a detergent and nuclease act up simultaneously, cell debris and nucleic acid components can be relatively rapidly and effectively removed, and the situation that the trenchancy of the cornea is degraded as protein components such as cornea collagen are damaged in long-term treatment of the detergent and other components can be avoided.

Description

technical field [0001] The invention belongs to the technical field of preparation of medical materials, and in particular relates to a preparation method of decellularized cornea. Background technique [0002] Corneal disease is the second leading cause of blindness in my country. There are about 4 million corneal blind patients in China, and about 100,000 new corneal blind patients are added every year. However, only about 5,000-8,000 corneal transplants can be performed each year in China. The serious shortage of corneal donors has led to the vast majority of corneal blind patients waiting in line in the dark, and some even missed the best opportunity for treatment, resulting in permanent blindness. Corneal transplantation is divided into penetrating keratoplasty, lamellar keratoplasty and corneal endothelial transplantation. Among them, lamellar keratoplasty has lower requirements for donor materials, and is suitable for all patients without obvious corneal endothelial ...

AI Technical Summary

Problems solved by technology

In recent years, with the extensive and in-depth research of biological tissue engineering, heterogeneous corneal stromal materials prepared by decellularization and virus inactivation have achieved preliminary therapeutic effects in animal level and clinical application, but corneal stromal materials after decellularization However, problems such as poor transparency, decreased resistance caused by partial mechanical structure damage, and gradual thinning of the thickness have not been well resolved. Therefore, the cornea using acellular matrix cannot replace the human cornea in clinical practice. There is a serious shortage of donor materials, so optimizing the process of removing corneal stromal cells to keep the transparent and mechanical structure of the decellularized cornea is a current research hotspot, and it is also the hope for the reconstruction of corneal blindness in my country.

[0003] Most of the corneal decellularization methods reported in the past use basic buffers, such as PBS, HBSS, cell culture fluid, etc. to swell the cornea, and the cells are broken and eluted by physical, chemical or biological methods. Short-term treatment cannot effectively remove the cells. Components generally require a decellularization process of 24-72 hours. When treated for a long time, the orderly collagen structure of the cornea will often be destroyed, accompanied by the loss of corneal transparency, and the optical effect is poor, which cannot meet the surgical requirements for clinical vision restoration.

Although the transparency can be restored after glycerin dehydration, it will gradually become thinner or opaque for a long time after transplantation, and the goal of long-term vision recovery cannot be achieved.