Cartilage derived collagen sponge scaffold and preparation method thereof
A technology of collagen sponge and cartilage, which is applied in the field of biomedical materials, can solve the problems of inhomogeneous cartilage scaffold materials, inconsistency in repairing defects, and difficult industrial standardized production, etc., and achieve the effect of good adhesion, good repair effect and low cost
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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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