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Cartilage derived collagen sponge scaffold and preparation method thereof

A technology of collagen sponge and cartilage, applied in the field of biomedical materials, can solve the problems of inhomogeneous cartilage scaffold materials, inconsistent repairing defect effects, difficult industrial standardized production, etc., and achieve the effect of favorable adhesion, good repairing effect and low cost

Active Publication Date: 2010-11-24
WEST CHINA HOSPITAL SICHUAN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0011] The cartilage scaffold materials disclosed in the prior art are all natural matrix materials obtained by degreasing, decalcifying, decellularizing and other methods of crushed cartilage particles, and composite materials obtained by adding collagen to this material, plus cartilage supply Due to body differences, the prepared cartilage scaffold materials are not uniform, have poor reproducibility, and are difficult to control the quality. The heterogeneity of scaffold materials leads to inconsistent defect repair effects in different researchers' animal experiments, which are not comparable and difficult to control. Large-scale clinical application, more difficult for large-scale industrial standardized production

Method used

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  • Cartilage derived collagen sponge scaffold and preparation method thereof
  • Cartilage derived collagen sponge scaffold and preparation method thereof
  • Cartilage derived collagen sponge scaffold and preparation method thereof

Examples

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Effect test

Embodiment 1

[0051] Embodiment 1: Preparation of cartilage-derived collagen sponge scaffold of the present invention

[0052] 1. Take fresh pig ear cartilage and pre-freeze it in a refrigerator at -70°C, freeze-dry it in a vacuum freeze dryer for 24 hours, grind it with a liquid nitrogen freezer pulverizer for 30 minutes, and take cartilage particles with a particle size of 90 μm or less through a No. 8 sieve;

[0053] 2. Mix cartilage microparticles with trypsin EDTA mixture (0.25% Trypsin+0.1% EDTA, TNE) according to volume ratio V / V 1:8~1:12, shake at 4°C for 12~30hr, centrifuge at 1000g for 10min and discard. After washing with PBS repeatedly for 3 times, the supernatant was discarded by centrifugation;

[0054] 3. Mix cartilage microparticles with 0.5mol / L HAc at a volume ratio of 1:10, add pepsin (wet weight ratio of pepsin and cartilage microparticles is 2:100), and continuously stir at 4°C for 12-24 hours.

[0055] 4. Centrifuge at 110g for 5min, take the supernatant, adjust the p...

Embodiment 2

[0064] Embodiment 2: Preparation of cartilage-derived collagen sponge scaffold of the present invention

[0065] 1. Take fresh pork rib cartilage and pre-freeze it in a -70°C refrigerator, freeze-dry it in a vacuum freeze dryer for 24 hours, grind it with a liquid nitrogen freezer for 30 minutes, and take cartilage particles with a particle size of 90 μm or less through a No. 8 sieve;

[0066] 2. Mix cartilage microparticles with trypsin EDTA mixture (0.25% Trypsin+0.1% EDTA, TNE) according to volume ratio V / V 1:8~1:12, shake at 4°C for 12~30hr, centrifuge at 1000g for 10min and discard. After washing with PBS repeatedly for 3 times, the supernatant was discarded by centrifugation;

[0067] 3. Mix the cartilage microparticles with 0.5 mol / L HAc at a volume ratio of 1:5, and add pepsin (the wet weight ratio of pepsin to cartilage microparticles is 3:100). Stirring was continued at 4°C for 24 hr.

[0068] 4. Centrifuge at 110g for 5min, take the supernatant, adjust the pH valu...

Embodiment 3

[0072] Embodiment 3: Preparation of cartilage-derived collagen sponge scaffold of the present invention

[0073] 1. Take fresh bovine ear cartilage and pre-freeze it in a refrigerator at -70°C, freeze-dry it in a vacuum freeze dryer for 24 hours, grind it with a liquid nitrogen freezer pulverizer for 30 minutes, and take cartilage particles with a particle size of 90 μm or less through a No. 8 sieve;

[0074] 2. Mix cartilage microparticles with trypsin EDTA mixture (0.25% Trypsin+0.1% EDTA, TNE) according to volume ratio V / V 1:8~1:12, shake at 4°C for 12~30hr, centrifuge at 1000g for 10min and discard. After washing with PBS repeatedly for 3 times, the supernatant was discarded by centrifugation;

[0075] 3. Mix the cartilage microparticles with 0.5 mol / L HAc at a volume ratio of 1:10, and add pepsin (wet weight ratio of pepsin to cartilage microparticles is 1:100). Stirring was continued at 4°C for 24 hr.

[0076] 4. Centrifuge at 110g for 5min, take the supernatant, adjus...

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Abstract

The invention discloses a method for preparing a cartilage derived collagen sponge scaffold and a collagen sponge scaffold prepared by the method. The method comprises the following steps of crushing, removing cells, removing telopeptide, dissolving and crosslinking, and the obtained collagen sponge scaffold has good biocompatibility and extremely low antigenicity; and a homogenized sponge scaffold can be prepared from cartilages of a plurality of individuals from the conspecific animals, has controllable pore diameter and porosity, can provide appropriate growth and propagation environment for cartilage cells and bone mesenchymal stem cells, particularly has good effect of repairing cartilage defect parts, and is more favorable for standardized production and clinical use.

Description

technical field [0001] The invention relates to a scaffold, in particular to a method for preparing a cartilage-derived collagen sponge scaffold and a product prepared by the method, belonging to the field of biomedical materials. Background technique [0002] After cartilage is damaged, it lacks the ability to effectively repair itself. At present, there are not many methods for clinical treatment of articular cartilage injury, and the effect is not ideal. Tissue engineering technology may become a new means of repairing articular cartilage defects. Natural biological scaffold materials have the advantages of cell signal recognition, promotion of cell adhesion, proliferation and differentiation, good biocompatibility and good biodegradability. Among them, the natural acellular bioscaffold material mainly utilizes the acellular matrix material obtained by chemical treatment of the same or heterogeneous organ / tissue. Extracellular matrix (Extracellular matrix, ECM) is secr...

Claims

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Application Information

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IPC IPC(8): A61L27/24A61L27/56
Inventor 解慧琪罗静聪杨志明
Owner WEST CHINA HOSPITAL SICHUAN UNIV
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