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Accellular epimatrix and preparation method and application thereof

A decellularization and extramatrix technology, applied in the field of medical materials, can solve problems such as inability to adapt to repair, inability to completely remove animal-derived DNA components, and damage to active components.

Inactive Publication Date: 2018-07-10
SHENZHEN LANDO BIOMATERIALS
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

There are many methods for preparing acellular extracellular matrix, but most of them cannot completely remove all animal-derived DNA components, and it takes a long time to use a variety of organic and surfactant solvents. On the one hand, the active components in the acellular matrix material On the other hand, harmful solvent residues can cause cytotoxicity, irritation and immune response, thereby affecting the tissue repair effect
And most of the acellular extracellular matrix is ​​not hydrophilic and sticking, the water absorption rate is slow, the film is not thin and soft enough, so it cannot adapt to the repair of various surface contour tissues
[0005] The orderly structure of the collagen fibers of the acellular extracellular matrix prepared by the existing method for preparing the extracellular matrix is ​​destroyed, thereby affecting the subsequent application

Method used

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  • Accellular epimatrix and preparation method and application thereof
  • Accellular epimatrix and preparation method and application thereof
  • Accellular epimatrix and preparation method and application thereof

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[0038] like figure 1 As shown, the present invention discloses a preparation method of decellularized extracellular matrix in one embodiment, comprising the following steps:

[0039] S10. Obtaining tissue precursors after preprocessing mammalian tissues and organs.

[0040] Pretreatment of mammalian tissues and organs is as follows: after preliminary removal of fat, muscle and other appendages from the mammalian tissues and organs by mechanical methods, they are cleaned with pure water (preferably 3 to 5 times). Of these, other organs attached indicate organs other than the pericardium, pleura, diaphragm, peritoneum, and small intestinal submucosa.

[0041] Preferably, the tissues and organs of mammals are those of pigs, cattle or horses.

[0042] Preferably, mammalian tissues and organs include mammalian pericardium, pleura, diaphragm, peritoneum and small intestinal submucosa.

[0043] Particularly preferably, the mammalian tissue is pig peritoneum.

[0044] The tissue p...

Embodiment 1

[0074] Take fresh pig peritoneum tissue, remove fat, muscle and other attached organs from the pig peritoneum by mechanical method, and remove other attached components to make pig peritoneum with a thickness of 0.3mm ~ 1.0mm, and wash it with pure water 4 times , stored at -20°C for later use.

[0075] Soak the pretreated pig peritoneum successively in 2wt% NaOH solution for 12 hours, wash it with pure water for 3 to 5 times, then treat it with 0.5wt% HCl solution for 2 hours, and wash it with pure water for 4 hours. to remove most of the DNA from the animal tissue.

[0076] The pig peritoneum after DNA removal treatment was soaked in 2wt% hydrogen peroxide solution for 3 hours to carry out virus inactivation treatment.

[0077] The inactivated pig peritoneum was soaked in 0.25wt% neutral protease solution for 24 hours, rinsed with pure water and soaked in 1M hypertonic saline for 24 hours to perform decellularization treatment.

[0078] The pig peritoneum was put into n-he...

Embodiment 2

[0090] Take fresh bovine peritoneum tissue, use mechanical method to preliminarily remove fat, muscle and other attached organs, and remove other attached components to make bovine peritoneum with a thickness of 0.3mm ~ 1.0mm, wash it with pure water for 5 times , stored at -20°C for later use.

[0091] Soak the pretreated bovine peritoneum successively in 2wt% NaOH solution for 10 hours, wash 5 times with pure water after treatment, then treat with 0.25wt% HCl solution for 4 hours, wash 5 times with pure water after treatment, to remove most of the DNA from animal tissues.

[0092] The bovine peritoneum after the DNA removal treatment was soaked in a mixed solution of 1 wt% peracetic acid and 50 wt% ethanol for 2 hours to perform virus inactivation treatment.

[0093] The inactivated bovine peritoneum was soaked in 0.25wt% trypsin solution for 12h, rinsed with pure water and soaked in 2M hypertonic saline for 12h for decellularization.

[0094] The bovine peritoneum was put...

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Abstract

The invention discloses a preparation method of an accellular epimatrix, the accellular epimatrix prepared by the preparation method and application of the accellular epimatrix in the field of preparation of medical instruments. The preparation method of the accellular epimatrix comprises the following steps: carrying out pretreatment on tissues and organs of mammals to obtain an analogy tissue precursor; separately carrying out virus inactivation treatment, DNA eliminating treatment, accellular treatment and defatting treatment on the analogy tissue precursor to obtain an accellular epimatrixprecursor; and successively carrying out gradient dewatering treatment and crosslinking treatment on the accellular epimatrix precursor, and finally shaping and drying to obtain the accellular epimatrix. By the gradient dewatering treatment, the accellular epimatrix precursor is soaked in organic matter solutions of which the concentrations are successively increased successively. According to the preparation method of the accellular epimatrix, the tissue precursor is treated through gradient dewatering, and thus, the orderly structure of collagenous fibers of the finally prepared accellularepimatrix is not damaged.

Description

technical field [0001] The invention relates to the field of medical materials, in particular to a decellularized extracellular matrix and its preparation method and application. Background technique [0002] The main development direction is the extracellular matrix material based on the principle of tissue engineering, which uses animal tissue as raw material. The extracellular matrix is ​​a complex and organic three-dimensional overall structure constructed from a variety of macromolecular substances such as collagen, non-collagenous glycoproteins, and elastin. It provides a suitable place and microenvironment for the survival and activities of various cells, and regulates tissues and organs. Function. Therefore, extracellular matrix, as an ideal tissue repair material, has been widely used clinically, including extracellular matrix materials such as pericardium, pleura, diaphragm, peritoneum and small intestinal submucosa. [0003] The sources of autologous and allogen...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61L27/36A61L27/50
CPCA61L27/3633A61L27/3687A61L27/50A61L2430/40
Inventor 李丽花钟梅玲朱勇军朱光林谭荣伟佘振定
Owner SHENZHEN LANDO BIOMATERIALS
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