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Preparation method and application of bioengineering decellularized eye conjunctiva

A decellularization and conjunctival technology, which is applied in the field of preparation of bioengineered medical materials, can solve the problems of loss of basic biological function of decellularized tissue and damage to the internal structure of tissue.

Active Publication Date: 2016-07-20
拜欧迪赛尔成都生物科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

These decellularization methods often lead to the destruction of the internal structure of the tissue and the loss of the original basic biological functions of the decellularized tissue

Method used

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  • Preparation method and application of bioengineering decellularized eye conjunctiva
  • Preparation method and application of bioengineering decellularized eye conjunctiva
  • Preparation method and application of bioengineering decellularized eye conjunctiva

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0030] Conjunctival tissue obtained by low-temperature fixer and trypan blue staining. By using a low-temperature fixer, the conjunctival tissue can be flattened and fixed on a low-temperature ice plate, and then the conjunctiva and subconjunctival tissue can be clearly distinguished by conventional trypan blue staining, and finally the epithelial layer is smooth and the tissue is flat. Conjunctival material.

[0031] 1. Protect the conjunctival tissue with protective solution throughout the whole process. The formula of the protective solution is: RPMI-1640 medium, add chondroitin sulfate 25g / L, low molecular dextran 10g / L, tobramycin 2.5mg / L, adjust pH value to 7.2, the osmotic pressure is 350mOsm;

[0032] 2. Seal the fresh conjunctiva after the subconjunctival tissue has been removed in a plastic bag filled with protective solution;

[0033] 3. Under the condition of 300MPa high static pressure, treat twice, each time is 2 minutes;

[0034] 4. After taking out the conju...

Embodiment 2

[0038] Conjunctival tissue obtained by low-temperature fixer and trypan blue staining. By using a low-temperature fixer, the conjunctival tissue can be flattened and fixed on a low-temperature ice plate, and then the conjunctiva and subconjunctival tissue can be clearly distinguished by conventional trypan blue staining, and finally the epithelial layer is smooth and the tissue is flat. Conjunctival material.

[0039] 1. Use the protective solution to protect the conjunctival tissue throughout the whole process. The formula of the protective solution is:

[0040] RPMI-1640 medium, add chondroitin sulfate 40g / L, low molecular weight dextran 20g / L, tobramycin 2.5mg / L, adjust the pH value to 7.2, and the osmotic pressure is 350mOsm;

[0041] 2. Seal the fresh conjunctiva after the subconjunctival tissue has been removed in a plastic bag filled with protective solution;

[0042] 3. Under the condition of 250MPa high static pressure, treat 5 times, each time is 1 minute;

[0043...

Embodiment 3

[0047] Conjunctival tissue obtained by low-temperature fixer and trypan blue staining. By using a low-temperature fixer, the conjunctival tissue can be flattened and fixed on a low-temperature ice plate, and then the conjunctiva and subconjunctival tissue can be clearly distinguished by conventional trypan blue staining, and finally the epithelial layer is smooth and the tissue is flat. Conjunctival material.

[0048] 1. Protect the conjunctival tissue with protective solution throughout the whole process. The formula of the protective solution is: RPMI-1640 medium, add chondroitin sulfate 50g / L, low molecular weight dextran 5g / L, tobramycin 2.5mg / L, adjust pH value to 7.2, the osmotic pressure is 380mOsm;

[0049] 2. Seal the conjunctiva after the subconjunctival tissue has been removed in a plastic bag filled with protective solution;

[0050] 3. Under the condition of 200MPa high static pressure, treat 3 times, each time is 2 minutes;

[0051] 4. After taking out the con...

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Abstract

The invention provides a preparation method of an efficient decellularized eye conjunctiva capable of keeping functions and characteristics of the eye conjunctiva. The method comprises the following steps of: acquiring a fresh eye conjunctiva which only contains less subconjunctival tissues; utilizing a mixed protective solution containing one or more conjunctiva structure protective agents during a whole eye conjunctiva decellularization process; pre-dissociating cells in the conjunctiva tissues by use of a high hydrostatic pressure technology; digesting residual cell nucleus by use of a composite nuclease method; removing loose smudge cell fragments by use of a detergent; and rinsing by use of the mixed protective solution. The decellularized eye conjunctiva has the advantage that complete structure, special fiber structure arrangement and biological characteristics of a conjunctival epithelium fibrous layer are maintained on the basis of ensuring thorough removal of cell nucleus.

Description

technical field [0001] The invention belongs to the field of preparation of bioengineering medical materials, in particular to a preparation method of decellularized conjunctiva. Background technique [0002] Conjunctival defects and related diseases are the main blinding eye diseases in my country. The normal environment and function of the conjunctiva is an indispensable condition for the normal visual function of the eyeball. Since our country is a big manufacturing and agricultural country, it is not uncommon to see eye diseases caused by conjunctival defects caused by physical or chemical eye trauma during production and agricultural work. In addition, in clinical ophthalmology, there are often conjunctival-related cases such as the resection of pterygium, if the defective conjunctival part is not handled properly, it will easily lead to the recurrence of pterygium. These eye diseases are very troublesome, and there is currently no good treatment. [0003] The curren...

Claims

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Application Information

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IPC IPC(8): A61L27/36A61L27/50
CPCA61L27/3604A61L27/50A61L2430/16A61L2430/40
Inventor 史真史伟云
Owner 拜欧迪赛尔成都生物科技有限公司