A bioactive high-purity collagen sponge and its preparation method
A collagen sponge and trypsin technology, applied in the field of medical biomaterials, can solve the problems of lack of high-purity, biologically active structure, etc.
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Embodiment 1
[0109] Fresh bovine Achilles tendons aged 18-24 months were taken, the fascia was removed, washed with purified water three times for 4 minutes each time, and then frozen in bundles in a low-temperature refrigerator at a temperature of -20 to -40°C. Freezing time ≥ 12h, and avoid repeated freezing and thawing. Cut the frozen Achilles tendon into 0.5mm slices with a microtome. Take by weighing 40g of bovine Achilles tendon thin slices, soak 3 times with 75% (V / V) ethanol aqueous solution of 1% (m / V) sodium bicarbonate as degreasing liquid, remove fat after soaking time is 20min, rinse 3 times with purified water , the time is 4 minutes until the pH is close to that of purified water, which is neutral.
[0110] Take the above treated Achilles tendon slices and disperse them in 2L of purified water, adjust the pH to 8.0, add trypsin with a final concentration of 0.1%, react at a reaction temperature of 37°C for 2 hours, rinse with purified water for 3 times for 4 minutes until t...
Embodiment 2
[0119] Take fresh Achilles tendon of 18-24 months old cattle, remove the fascia, rinse with purified water 5 times for 5 minutes, and then freeze them in bundles. The freezing temperature is -20 to -40°C, and the freezing time is for 12 hours, and avoid repeated freezing and thawing. Cut the frozen Achilles tendon bundle into 0.5mm thin slices with a microtome, weigh 40 g of the bovine Achilles tendon, soak it with 75% (V / V) ethanol aqueous solution of 1% (m / V) sodium bicarbonate as degreasing liquid 5 times, after soaking for 30 minutes to remove fat, rinse with purified water 5 times, each time for 5 minutes until the pH is close to that of purified water and becomes neutral.
[0120] Take the above treated Achilles tendon slices and disperse them in 1L of purified water, adjust the pH to 8.2, add trypsin with a final concentration of 0.1%, react at a reaction temperature of 40°C for 2 hours, rinse with purified water 5 times, each time for 5 minutes to reach the pH Close t...
Embodiment 3
[0129] Take fresh Achilles tendon of 18-24 months old cattle, remove the fascia, rinse with purified water 4 times for 3 minutes, and then freeze them in bundles. The freezing temperature is -20 to -40°C, and the freezing time is for 15 hours, and avoid repeated freezing and thawing. With slicer, frozen Achilles tendon bundle is cut into thin slices of 5 mm, and 40 g of bovine Achilles tendon is taken, soaked with 90% (V / V) ethanol aqueous solution of 5% (m / V) sodium bicarbonate as degreasing liquid for 2 After soaking for 15 minutes to remove fat, rinse with purified water for 4 times, each time for 10 minutes until the pH is close to that of purified water and becomes neutral.
[0130] Take the above treated Achilles tendon slices and disperse them in 400mL of purified water, adjust the pH to 8.5, add trypsin with a final concentration of 0.025%, react at a reaction temperature of 40°C for 2h, wash with purified water 4 times, each time for 10min to pH Close to purified wat...
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