A preparation method of decellularized nucleus pulposus material derived from natural tissue

A decellularization and tissue technology, applied in the field of preparation of decellularized nucleus pulposus materials, to achieve good biocompatibility, low immune rejection, and promote the regeneration of degenerated nucleus pulposus

Active Publication Date: 2021-04-06
ZHEJIANG DISAI BIOTECHNOLOGY CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, there is no report on the decellularized scaffold of nucleus pulposus with excellent performance

Method used

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  • A preparation method of decellularized nucleus pulposus material derived from natural tissue
  • A preparation method of decellularized nucleus pulposus material derived from natural tissue
  • A preparation method of decellularized nucleus pulposus material derived from natural tissue

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0035]Example 1: Preparation method and regenerative repair of nucleus marrow deodorcular materials from natural tissues

[0036]Preparation of fibrus

[0037](1) Take the material: Take the slaughter of the pig's chest 5 to the waist 6 spine, separated from the spine, then cut the fiber ring to remove the nucleus, the medullary is about 1.5 * 1.5 * 1 cm. The pulp core was rinsed 3 times with a PBS buffer containing 10K IU / mL protease inhibitor, 30 min, remove blood, attached fat fragments and other impurities.

[0038](2) The decellular steps are as follows:

[0039]1, in the PBS buffer containing 20K IU / mL protease inhibitor, a low temperature (4 ° C) shaker is oscillated for 24 hours, and then the liquid nitrogen is frozen three cycles (-80 ° C / 37 ° C);

[0040]2, in the PBS buffer containing 1 M NaCl, 1: 1, penicillin and streptomycin (20 kiu / ml, 20 g / ml) mixed antibacterial solution, low temperature (4 ° C) shaker 100 rpm shock 24 hours;

[0041]3, in PBS buffer having a concentration ...

Embodiment 2

[0053]Example 2: Preparation and Rehabilitation Rehabilitation of Determination Melodeling Materials

[0054](1) Take the material: Take the slaughter of the pig's chest 5 to the lumbar 6 spine, separated the discharged discharge with a knife, then cut the fiber ring to remove the nucleus, the nucleus is about 1 * 1 * 0.6 cm. The pulp core was rinsed 3 times with a PBS buffer containing 10K IU / mL protease inhibitor, 30 min, remove blood, attached fat fragments and other impurities.

[0055](2) The decellular steps are as follows:

[0056]1, in the PBS buffer containing 15K iU / mL protease inhibitor, a low temperature (4 ° C) shaker is oscillated for 36 hours, then freeze three circulation (-80 ° C / 37 ° C) with liquid nitrogen.

[0057]2, in the PBS buffer containing 1.2 M NaCl, 1: 1, penicillin and streptomycin (20 kiu / ml, 20 g / ml) mixed antibacterial solution, low temperature (4 ° C) shaker 100 rpm shock for 36 hours;

[0058]3, in PBS buffer having a concentration of 0.08% of Triton X-1...

Embodiment 3

[0061]Example 3: Preparation and Regeneration Rehabilitation of Deparaginucleus Materials

[0062](1) Take the material: Take the slaughter of 12h with the bottom of the sheep 5 to the waist 6 spine, separated the intervertebral disc with a knife, then cut the fiber ring to remove the nucleus, the medullary is about 0.6 * 0.6 * 0.6 cm. The pulp core was rinsed 3 times with a PBS buffer containing 10K IU / mL protease inhibitor, 30 min, remove blood, attached fat fragments and other impurities.

[0063](2) The decellular steps are as follows:

[0064]1, in the PBS buffer containing 20K IU / mL protease inhibitor, a low temperature (4 ° C) shaker oscillated for 48 hours, then freezing three cycles with liquid nitrogen (-80 ° C / 37 ° C);

[0065]2, in the PBS buffer containing 1.5 M NaCl, 1: 1 was added with penicillin and streptomycin (20 kiu / ml, 20g / ml) mixed antibacterial liquid, low temperature (4 ° C) shaker 100 rpm shock for 48 hours;

[0066]3, in the PBS buffer of Triton X-100 in concent...

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Abstract

The invention discloses a method for preparing a decellularized nucleus pulposus material derived from natural tissue. The nucleus pulposus tissue of any size in a mammal is passed through PBS containing protease inhibitors, PBS buffer containing NaCl, and PBS buffer containing Triton X. , PBS buffer containing SDS, and PBS buffer containing DNase to obtain decellularized nucleus pulposus material from natural tissue. The invention can completely decellularize the cells on the tissue with mild conditions, no ECM damage, and rapid stability. The obtained material not only imitates the natural nucleus pulposus structure to the greatest extent, better retains the extracellular matrix components, but also has the advantages of extremely low immunogenicity. It can be used to repair intervertebral disc injuries caused by various etiologies in clinical practice, Diseases associated with degeneration of the nucleus pulposus.

Description

Technical field[0001]The present invention is mainly used in the repair of intervertebral disc myelus nucleus and regenerative medical fields. It is specifically involved in the preparation method of a nuclear deodorcular material of a natural tissue source.Background technique[0002]Intervertebral disc degeneration is the main factor that leads to highlighted lumbar disc herniation, bringing huge burden to social security and economic. The surgery in the absence of lumbar disc herniation is mainly vertebral fusion, and does not restore the function of normal intervertebral disc. The organizational engineering will resume normal intervertebral disc physicism and mechanical functions, and provide a possible way for future intervertebral disc lesions. The histological characteristics of intervertebral disc degeneration is that the amount of marrow content is reduced, and the II collagen is increased and the type II collagen is reduced. At the same time, the glycosamine content is also ...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): A61L27/36A61L27/50
CPCA61L27/3633A61L27/3658A61L27/3687A61L27/50A61L2430/38A61L2430/40
Inventor 林贤丰许家齐方向前范顺武王晟毓
Owner ZHEJIANG DISAI BIOTECHNOLOGY CO LTD
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