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Lamellar corneal stroma bracket as well as preparation method and application thereof

A stromal scaffold and lamellar cornea technology, applied in medical science, prosthesis, etc., can solve the problems of large damage to collagen lamellar structure, disorderly arrangement of collagen fibers, uneven pores, etc., and achieve no cell residue and biocompatibility. And the effect of good biological safety and uniform pores

Active Publication Date: 2014-08-27
QINGDAO CHUNGHAO TISSUE ENG
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0006] The purpose of the present invention is to overcome the large damage to the collagen lamellar structure of the corneal stroma substitute prepared by the existing method, which makes the arrangement of collagen fibers messy, the pores are uneven, and the light transmittance is low, which cannot fully meet the clinical requirements. Deficiencies in transplantation requirements, providing a lamellar corneal stromal scaffold with neatly arranged collagen fibers, uniform and regular pores, and high light transmittance, as well as its preparation method and application

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  • Lamellar corneal stroma bracket as well as preparation method and application thereof
  • Lamellar corneal stroma bracket as well as preparation method and application thereof
  • Lamellar corneal stroma bracket as well as preparation method and application thereof

Examples

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Effect test

Embodiment 1

[0047] This example is used to illustrate the lamellar corneal stromal stent of the present invention and its preparation method.

[0048] (1) Take the pig eyeball, and soak the fresh pig eyeball with 75% alcohol by volume for 20 seconds, and directly cut out a corneal stroma sheet with a diameter of 9 mm and a thickness of 400 μm in an ultra-clean bench;

[0049] (2) Soak the cut corneal stroma sheet in triple distilled water for 10 hours;

[0050] (3) Place the hypoosmotically swollen corneal stroma sheet obtained in step (2) in a sterile cryopreservation tube, freeze and thaw repeatedly, the freezing time is 40 minutes, the freezing temperature is -80 ° C, the melting time is 20 minutes, and the melting temperature is 37°C, the number of cycles is 3 times;

[0051] (4) Put the corneal stromal slices that have been repeatedly frozen and thawed in Tris-Hcl buffer containing DNase and RNase for enzymatic digestion. The volume ratio of DNase to Tris-Hcl buffer is 1:1000, and R...

Embodiment 2

[0058] This example is used to illustrate the lamellar corneal stromal stent of the present invention and its preparation method.

[0059] (1) Take the bull's eyeball, soak the fresh bull's eyeball in 1×PBS solution containing tobramycin (the concentration of tobramycin is 40000U / L) for 30s, and cut it directly with a keratome in an ultra-clean bench Corneal stroma sheet with a diameter of 8 mm and a thickness of 200 μm;

[0060] (2) Place the cut corneal stroma piece in triple distilled water and soak for 8 hours;

[0061] (3) Place the hypotonic and swollen corneal stroma sheet obtained in step (2) in a sterile cryopreservation tube, freeze and thaw repeatedly, the freezing time is 30min, the freezing temperature is -200°C, the melting time is 30min, and the melting temperature is 40°C, the number of cycles is 2;

[0062] (4) Put the corneal stromal slices that have been repeatedly frozen and thawed in 1×PBS buffer containing DNase and RNase for enzymatic digestion. The vo...

Embodiment 3

[0069] This example is used to illustrate the lamellar corneal stromal stent of the present invention and its preparation method.

[0070] (1) Take the monkey eyeball, and soak the fresh monkey eyeball with 80% alcohol by volume for 10 seconds, and in the ultra-clean bench, directly cut out a corneal stroma sheet with a diameter of 10 mm and a thickness of 600 μm with a keratome;

[0071] (2) Place the cut corneal stroma piece in triple distilled water and soak for 12 hours;

[0072] (3) Place the hypotonic swollen corneal stroma sheet obtained in step (2) in a sterile cryopreservation tube, freeze and thaw repeatedly, the freezing time is 60 minutes, the freezing temperature is -50 ° C, the melting time is 15 minutes, and the melting temperature is 30°C, the number of cycles is 5 times;

[0073] (4) Put the corneal stromal piece that has been repeatedly frozen and thawed in Hanks buffer containing DNase and RNase for enzymatic digestion. The volume ratio of DNase and Hanks b...

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Abstract

The invention discloses a lamellar corneal stroma bracket as well as a preparation method and application thereof. The method comprises the following steps: carrying out low-permeability swelling, repetitive freeze-thawing, enzyme digesting, drying and sterilizing treatment on a corneal stroma sheet of a fresh animal eyeball cut down under a sterile condition, wherein the enzyme digesting adopts buffer liquor containing DNA (deoxyribonucleic acid) enzyme and RNA (ribonucleic acid) enzyme to treat, ultrasonic treatment is carried out after the enzyme digestion treatment, and then drying and sterilizing treatment is carried out. The invention further provides application of the lamellar corneal stroma bracket as a corneal stroma substrate. The method disclosed by the invention can be used for reducing damages on a corneal stroma collagen structure to the greatest extent; collagenous fiber arrangement is orderly, holes are uniform and regular, cell residues are avoided, the lamellar structure is kept complete, and the biocompatibility of the bracket material is improved.

Description

technical field [0001] The invention relates to the field of corneal stroma substitutes, in particular to a lamellar corneal stroma support and its preparation method and application. Background technique [0002] The cornea is located on the outermost surface of the eyeball and is in direct contact with the outside world. It is vulnerable to damage (physical, chemical) and infection. Once a lesion occurs, it will become cloudy and affect vision, or even cause blindness. Common corneal lesions include keratitis, corneal ulcer, keratoconus, keratomacia, and corneal degeneration. According to the statistics of the World Health Organization (WHO), there are nearly 50 million people with corneal disease blindness in the world at present, and there are about 4 million people in my country. Corneal disease has seriously affected the patients' normal life, work and study. They need family and social care, which increases the financial burden of the family and the burden of the who...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61L27/36A61L27/24
Inventor 李青王宝泉彭艳婷王红梅李海燕
Owner QINGDAO CHUNGHAO TISSUE ENG
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