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Decellularization method for preparing extracellular matrix (ECM) support material

A scaffold material and extramatrix technology, used in medical science, tissue regeneration, prosthesis, etc., can solve the problems of sodium lauryl sulfate toxicity, many methods for removing detergents, damage to matrix structure, etc., and achieve a good ecological environment. Safety and compatibility, enhanced decellularization effect, reduced irritation effect

Active Publication Date: 2014-01-29
SHANGHAI MICROPORT CARDIOFLOW MEDTECH CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] Chinese Patent Application No. 201110134511.9 relates to a preparation method of the same kind of acellular dermal matrix, which uses sodium dodecyl sulfate for decellularization treatment. Although the time is short, sodium dodecyl sulfate has great damage to the matrix structure. Moreover, sodium lauryl sulfate has obvious toxicity, which is not conducive to implantation in the body.
To avoid this toxic reaction, it is necessary to remove the detergent or replace it with a new non-toxic reagent. There are many ways to remove the detergent and the process is cumbersome and incomplete. Organ cell replacement reagents

Method used

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  • Decellularization method for preparing extracellular matrix (ECM) support material
  • Decellularization method for preparing extracellular matrix (ECM) support material
  • Decellularization method for preparing extracellular matrix (ECM) support material

Examples

Experimental program
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Effect test

Embodiment 1

[0044] 1) Material preparation: Pig pericardium was obtained from a local slaughterhouse. After slaughtering a healthy adult pig, the heart was taken out, and the pericardium was cut. The warm ischemia time was less than 2 hours. Phosphate buffered solution (PBS) was repeatedly washed to remove blood clots, then placed in the preservation solution, and brought back to the laboratory, the peripheral fat was removed under aseptic conditions, and the undamaged and uniform thickness of the anterior wall was trimmed into small pieces of 3cm×4cm. Rinse thoroughly with PBS solution, and store in sterile PBS solution containing antibiotics at 4°C.

[0045] 2) Decellularization of porcine pericardium: Put 6 pieces of 3cm×4cm pericardium into 6 bottles of 1% OGP (Aladdin Reagent (Shanghai) Co., Ltd.) 10mM Tris-HCl buffer solution for shaking digestion for 24h, 4°C , 150rpm / min, each bottle of solution contains 1% streptomycin sulfate (Aladdin Reagent (Shanghai) Co., Ltd., batch number: ...

Embodiment 2

[0060] After preparing the porcine pericardium tissue according to the method of Example 1, put 6 pieces of 3cm×4cm pericardium into 6 bottles of 10mM Tris-HCl buffer solution of 2% OGP for shaking digestion for 16h, 4°C, 150rpm / min, each bottle The solution contained 1% double antibody solution of streptomycin sulfate and penicillin sodium. Afterwards, wash in sterile PBS (pH7.30), 15 min / time, 5 times; after washing, place the pericardium in a nuclease solution at 37° C. for 24 h with shaking and digestion. Nuclease solution: 2.5KU / ml DNaseⅠ, 7.5KU / ml RNase, 0.15M NaCl, 2mMMgCl 2 (H 2 O) 6 and 1% double antibody sterile 50mM Tris-HCl buffer (pH7.60). Shake and wash in sterile double-antibody-containing PBS buffer (pH 7.30) for 24 hours at a rotational speed of 150 rpm. After washing, the pericardium is sequentially stored at 4° C. in the double-antibody-containing sterile PBS solution.

Embodiment 3

[0062]After the porcine pericardium tissue was prepared according to the method in Example 1, 6 pericardiums of 3 cm × 4 cm were put into 10 mM Tris-HCl of 6 bottles of 1% dodecyl glucopyranoside (Aladdin Reagent (Shanghai) Co., Ltd.) respectively. Perform shaking digestion in the buffer solution for 24 hours, 4°C, 150rpm / min, and each bottle of solution contains a double antibody solution of 1% streptomycin sulfate and penicillin sodium. Afterwards, wash in sterile PBS (pH7.30), 15 min / time, 5 times; after washing, place the pericardium in a nuclease solution at 37° C. for 24 h with shaking and digestion. Nuclease solution: 2.5KU / ml DNaseⅠ, 7.5KU / ml RNase, 0.15M NaCl, 2mM MgCl 2 (H 2 O) 6 and 1% double antibody sterile 20mM Tris-HCl buffer (pH7.60). Shake and wash in sterile double-antibody-containing PBS buffer (pH 7.30) for 24 hours at a rotational speed of 150 rpm. After washing, the pericardium is sequentially stored at 4° C. in the double-antibody-containing sterile P...

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Abstract

The invention relates to the field of tissue engineering materials, and in particular relates to a decellularization method for preparing an extracellular matrix (ECM) support material. By using green surfactant glucopyranoside for decellularization treatment, cells can be completely removed, and an ultra microstructure and compositions of an ECM support are maintained.

Description

technical field [0001] The invention relates to the field of tissue engineering materials. Specifically, the present invention relates to a decellularized method for preparing extracellular matrix scaffold materials. Background technique [0002] Tissues are composed of cells and extracellular matrix (ECM), the cellular components of which are recognized by the host as antigens, which will trigger inflammation or immune rejection. While the ECM is a complex of structural and functional proteins, this component is generally protected and well tolerated across species. Bioscaffold materials composed of ECM are widely used in the field of surgical reconstruction and regenerative medicine of tissues and organs, such as heart valves, blood vessels, skin, nerves, tendons, and small intestinal submucosa. ECM is a secreted product of cells in tissues and organs. It has a dynamic interaction with these cells, can reflect changes in the microenvironment in time, and plays an importa...

Claims

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Application Information

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IPC IPC(8): A61L27/36
CPCA61L2430/40A61L27/3629A61L27/3625A61L27/362A61L27/3687
Inventor 董教明李雨陈大凯陈叶萌王静穆元姗房圆陈国明张晓怡乐承筠罗七一
Owner SHANGHAI MICROPORT CARDIOFLOW MEDTECH CO LTD
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