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Highly-transparent acellular corneal stroma and preparation method thereof

A corneal stroma and decellularization technology, applied in the field of medical devices, can solve the problems of high cytotoxicity of materials, poor effect of cell migration and reconstruction, and reduced transparency, so as to avoid damage to collagen bundles and natural three-dimensional microstructure, and ensure the arrangement of collagen structures , reducing the effect of the immune response

Active Publication Date: 2020-09-22
SHAANXI BOYU REGENERATIVE MEDICINE CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, decellularization technology mainly focuses on physical, chemical and enzymatic decellularization methods. Freezing and thawing will cause collagen fibers to aggregate and break, and decellularization is not complete. High static pressure will lead to an increase in the melting point of water, and formation of The ice crystals disrupt the three-dimensional structure of collagen
It is easy to decellularize by enzymatic method, but the collagen will be degraded, and the enzyme is not easy to inactivate, resulting in disordered fiber results or damage to the three-dimensional microstructure, and reduced transparency; decellularized by detergent, which is not easy to remove, high reagent residue, and material cytotoxicity High, does not meet the material compatibility requirements, implanted in the body, the remodeling effect of its own cell migration is poor or inhibited

Method used

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  • Highly-transparent acellular corneal stroma and preparation method thereof
  • Highly-transparent acellular corneal stroma and preparation method thereof
  • Highly-transparent acellular corneal stroma and preparation method thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0038] The preparation method of the decellularized corneal stroma of the present embodiment comprises the following steps:

[0039] Step 1. Rinse the extracted animal eyeballs with normal saline, then place them in a sodium hypochlorite solution for virus inactivation, and finally wash and remove the sodium hypochlorite with normal saline; the mass concentration of the sodium hypochlorite solution is 0.1%, and the solvent for the sodium hypochlorite solution It is normal saline, and the time for virus inactivation treatment is 30 minutes;

[0040] Step 2, cutting the cornea from the animal eyeball after cleaning and virus inactivation in step 1, the cut cornea has a 3 mm wide sclera, and then cleaning the cut cornea with normal saline;

[0041] Step 3, place the cleaned cornea in step 2 in a hypotonic solution, and after ultrasonic treatment for 60s, transfer to a shaker for 0.5h; the hypotonic solution is normal saline, the frequency of ultrasonic treatment is 50Hz, and the ...

Embodiment 2

[0048] The present embodiment is the same as Example 1, wherein the difference is: the solvent of the sodium hypochlorite solution described in step 1 is a phosphate buffered saline solution with a pH value of 6 to 8 or a Tris buffered saline solution with a pH value of 6 to 8; The hypotonic solution described in the third is a phosphate buffer solution with a pH value of 6 to 8 or a Tris buffered saline solution with a pH value of 6 to 8; the solvent of the transparent solution described in step 5 is phosphoric acid with a pH value of 6 to 8 Salt buffer solution or Tris buffered saline solution with a pH value of 6-8, the liquid dehydrating agent is ethanol or butanediol, or two or three of glycerin, ethanol and butanediol, the solvent of the liquid dehydrating agent It is a phosphate buffer solution with a pH value of 6 to 8 or a Tris buffered saline solution with a pH value of 6 to 8; the sterile preservation solution described in step 7 is butylene glycol, propylene glycol ...

Embodiment 3

[0050] The preparation method of the decellularized corneal stroma of the present embodiment comprises the following steps:

[0051] Step 1. Rinse the extracted animal eyeballs with a phosphate buffer solution with a pH value of 6, then place them in a sodium hypochlorite solution for virus inactivation, and finally wash and remove the sodium hypochlorite with a phosphate buffer solution with a pH value of 6; The mass concentration of described sodium hypochlorite solution is 0.02%, and the solvent of sodium hypochlorite solution is the phosphate buffer solution that pH value is 6, and the time of virus inactivation treatment is 60min;

[0052] Step 2, cutting the cornea from the animal eyeball after cleaning and virus inactivation in step 1, the cut cornea has a 4mm wide sclera, and then cleaning the cut cornea;

[0053] Step 3, place the cleaned cornea in step 2 in a hypotonic solution, and after ultrasonic treatment for 30 s, transfer it to a shaker for 2 h; the hypotonic s...

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Abstract

The invention discloses a preparation method of a highly-transparent acellular corneal stroma. The preparation method comprises the following steps: 1, cleaning and virus inactivation treatment are carried out on a picked animal eyeball; 2, a cornea is shorn, and then the shorn cornea is cleaned; 3, the cornea is put into a hypotonic solution, subjected to ultrasonic treatment and then transferredinto a shaking table for shaking treatment; 4, the cornea is repeatedly treated 3-4 times; 5, hypertonic and hypotonic solution treatment is carried out, after every 1-2 times of hypertonic and hypotonic solution treatment, the cornea is put into a transparent solution to be treated for 0.5-3 h, and after the last time of hypertonic and hypotonic solution treatment, the cornea is dehydrated; 6, the dehydrated cornea is cut; and 7, the cut cornea is sterilized and then preserved. According to the method, the corneal stroma can be effectively prevented from absorbing water and swelling, collagen bundles and a natural three-dimensional microstructure are prevented from being damaged, therefore it is guaranteed that collagen structures of a Bowman's membrane and a stroma layer are arranged inorder, and the prepared acellular corneal stroma has the optical performance and strength similar to those of a fresh cornea.

Description

technical field [0001] The invention belongs to the technical field of medical devices, and in particular relates to a highly transparent decellularized corneal stroma and a preparation method thereof. Background technique [0002] Corneal disease is the second leading cause of blindness after cataracts. Because the cornea is in direct contact with the external environment for a long time, it is extremely vulnerable to external mechanical damage, microbial infection, burns, etc., resulting in lesions in the cornea or part of the cornea. In mild cases, the transparency of the cornea decreases. It affects the normal life of the patient, and in severe cases, it will cause corneal opacity or ulceration, causing blindness or removal of the eyeball. According to WTO statistics, there are currently about 60 million blind patients with corneal disease in the world, and there are 4 million people in China, and the number of cases continues to increase by 100,000 every year. The vast ...

Claims

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Application Information

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IPC IPC(8): A61L27/36A61L27/50
CPCA61L27/3633A61L27/3687A61L27/3691A61L27/50A61L2430/40A61L2430/16
Inventor 樊杏利任珂星王二浩张立娟张勇杰
Owner SHAANXI BOYU REGENERATIVE MEDICINE CO LTD
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