Fish skin collagen support loading epidermal growth factors and preparation method thereof
A technology of epidermal growth factor and collagen scaffold, applied in the field of medical devices, can solve the problems of rapid diffusion of epidermal growth factor, virus contamination, loss of biological activity, etc. short effect
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Embodiment 1
[0022] Embodiment 1: the preparation of fish skin collagen scaffold
[0023] (1) Thaw the fish skin and cut it into pieces;
[0024] (2) After washing with distilled water, add a mixed solution of hydrogen peroxide with a volume fraction of 1% and 0.01 mol / L sodium hydroxide that is 20 times its mass, stir mechanically for 24 hours, and replace the alkaline solution every 8 hours;
[0025] (3) Add isopropanol solution with a volume fraction of 10%, mechanically stir for 4 h, and wash with purified water;
[0026] (4) Add 2.5% sodium chloride solution, stir mechanically for 12 h, and wash with purified water;
[0027] (5) Add 10 volumes of purified water, adjust pH=2.0-3.0 with hydrochloric acid, add pepsin with 2.5% fish skin mass, enzymolyze at 4-8°C for 16-24 h, centrifuge at 8000-20000 rpm for 20 min, take Serum;
[0028] (6) Add a certain amount of sodium chloride to the above supernatant to a final salt concentration of 0.9mol / L, salt out, centrifuge at 8000 rpm for 20...
Embodiment 2
[0031] Example 2: Preparation of Nanoparticles Loaded with Epidermal Growth Factor
[0032] (1) Dissolve chitosan in 1% acetic acid, dialyze for 4 days, and freeze-dry to obtain purified chitosan;
[0033] (2) Dissolve 0.2 g chitosan in 5 ml 1% acetic acid, adjust the pH value to 6.0 with 0.1mol / L NaOH, and set the volume to 100 ml to obtain a chitosan solution with a concentration of 2 mg / ml;
[0034] (3) Prepare a heparin solution with a concentration of 1 mg / ml, and use this solution as the mother solution to prepare a heparin mixture solution containing epidermal growth factor 10500 IU / ml;
[0035] (4) At 4°C, under electric stirring, drop 2ml of the 1mg / mL above heparin mixture into 5ml of the 2 mg / ml chitosan solution drop by drop at a rate of 10-40 drops per minute to obtain dispersion Uniformly loaded nanoparticles of epidermal growth factor.
Embodiment 3
[0036] Example 3: Preparation of fish skin collagen scaffold loaded with epidermal growth factor
[0037] The fish skin collagen scaffold prepared in Example 1 was infiltrated with an aqueous solution of nanoparticles loaded with epidermal growth factor at a ratio of 2 ml / cm 2; Store at -20°C after infiltration. Vacuum freeze-drying, that is.
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