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Novel chondrocyte epimatrix membrane and preparation method thereof

A chondrocyte and acellular matrix technology, applied in the field of cartilage extracellular matrix membrane and its preparation, can solve the problem of inability to biomimetic cartilage extracellular matrix, and achieve the effect of reducing cytotoxicity and improving biological activity

Inactive Publication Date: 2014-02-26
成都军区昆明总医院
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0006] The present invention intends to overcome the disadvantage that the collagen film or biological film constructed by type I / III collagen and other polymers cannot mimic the extracellular matrix of cartilage, and proposes a novel cartilage extracellular matrix membrane and its preparation method. The extracellular matrix membrane It can be used to repair cartilage damage and prevent tissue adhesion such as tendon

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0031] Example 1: Preparation of Cartilage Acellular Matrix Slurry

[0032] Cut the hyaline cartilage of fresh cadaveric knee joints with a scalpel under the condition of a sterile ultra-clean bench, rinse with ice PBS solution at 4°C for 3 times, place the cartilage tissue in 10 mM Tris-HCl solution containing protease inhibitors, and clean it in a centrifuge at 3000 rpm After 3 times, place in 10 mM Tris-HCl solution containing protease inhibitors (1 mmol / l phenylmethylsulfonyl fluoride and 1% ethylenediaminetetraacetic acid) for later use.

[0033] The cleaned cartilage tissue was placed in ice Tris-HCl (10mM) solution containing protease inhibitors, the tissue was pulverized by a low-temperature tissue grinder, and the cartilage particles were screened through a filter with a diameter of 500nm-2mm.

[0034] Add 1% Triton X-100 Tris-HCl buffer (pH 7.5) to the centrifuged cartilage microparticles, which contains the aforementioned protease inhibitors, shake on a constant tem...

Embodiment 2

[0036] Example 2: Preparation of Cartilage Extracellular Matrix Membrane

[0037] Take 20ml of acellular matrix slurry with a concentration of 5%, put it into a glass container with an area of ​​9cm2, and place it in an environment at 4°C for 24 hours to properly accelerate the air circulation (wind speed 1-2m / s), and the acellular matrix slurry can be produced by itself Cross-linking, it can be prepared into a film after 48 hours. The scaffold was then placed under 254nm ultraviolet light at a distance of 10 cm for 12 hours, and the ultraviolet cross-linking increased the mechanical strength of the membrane. The extracellular matrix membrane can also be cross-linked using a chemical cross-linking agent (after immersing the scaffold in 40% ethanol solution for 30 min, use 5.5 mM NHS (N-hydroxysuccinimide) and 14 mM EDAC (carbodiimide) mixed aqueous solution, the cross-linked scaffold was washed at room temperature for 4 hours, washed 3 times with 0.01 mmol PBS, and the scaffo...

Embodiment 3

[0040] Inoculate the chondrocytes cultured in vitro at 1*106 / cm2 in the cartilage defect area, the cartilage defect area can be added with an extracellular matrix scaffold constructed from biosynthetic materials or biologically derived materials, and the extracellular matrix prepared according to Example 2 The cartilage defect area was sealed with a membrane and closed with fibrin gel or sutured with absorbable sutures. Hyaline cartilage repair could be observed at 8 weeks after operation.

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Abstract

The invention discloses a novel chondrocyte epimatrix membrane and a preparation method thereof. The membrane contains cell epimatrix ingredients which have bioactivity and cell activity, wherein the cell epimatrix comprises the following main ingredients in percentage by weight: 80 to 95 percent of bionic type II collagen in an acellular cartilage matrix and 10 to 20 percent of hyaluronic acid, 10 to 20 percent of aminopolysaccharide and the like which serve as a chondrocyte epimatrix; the chondrocyte epimatrix biological membrane prepared from the acellular cartilage matrix has the bionic chondrocyte epimatrix ingredients, is excellent in cell consistency, does not arouse immunological rejection of host cartilage tissue, can be used for repairing the damage of hyaline cartilage tissue of joints and reconstructing the hyaline cartilage tissue, and also can be used for a seed cell inoculating vector and a growth factor sustained-release vector; and the chondrocyte epimatrix contains blood vessel inhibiting factor ingredients, so the membrane also can be used for preventing the conglutination of tissue such as muscle tendons.

Description

technical field [0001] The invention relates to medical biomaterials, in particular, the invention is a novel cartilage extracellular matrix membrane and a preparation method thereof. Background technique [0002] The active use of articular cartilage is huge, and it is easy to cause damage in trauma and acute and chronic inflammation. It has been reported in the literature that the incidence of local cartilage damage in patients undergoing arthroscopic knee surgery is as high as 63%. The physiological mechanism of cartilage growth and metabolism is not completely clear, but it is currently believed that articular cartilage has no blood, no lymph, no nerves, contains only a single chondrocyte, has a high content of extracellular matrix, has few cells, and lacks local progenitor cells, resulting in self-repair of articular cartilage Poor ability. Late articular cartilage damage will eventually lead to osteocartilage stripping, joint exudation, and articular surface incomplet...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): A61L27/26A61L27/24A61L27/20A61L27/54
Inventor 谭洪波徐永清丁晶林月秋
Owner 成都军区昆明总医院
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