Collagen-chitosan tissue engineering scaffold compounded with tβ4 and its preparation method and application
A technology of tissue engineering scaffold and chitosan, which is applied in medical science, prosthesis, etc., can solve the problems of unfavorable skin refractory wound repair process, skin appendage regeneration and low efficiency, so as to promote rapid angiogenesis and re-epithelialization, Improving the efficiency of transplantation and promoting the effect of repair and regeneration
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[0020] In the first aspect, the present invention provides a preparation method of a collagen-chitosan tissue engineering scaffold compounded with Tβ4, the method comprising: preparing a collagen-chitosan porous scaffold, and soaking the collagen-chitosan porous scaffold in Tβ4 In the solution, a collagen-chitosan porous scaffold complexed with Tβ4 is obtained; then the collagen-chitosan porous scaffold complexed with Tβ4 is sequentially subjected to chemical cross-linking treatment and freeze-drying treatment.
[0021] In the preparation method of the present invention, the method for preparing the collagen-chitosan porous scaffold is not particularly limited, and may be various preparation methods commonly used in the art. But, the inventor of the present invention finds in research, when preparing collagen-chitosan porous support, control the specific concentration of collagen and chitosan and the pH value of collagen-chitosan solution in collagen-chitosan solution The cumu...
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[0038] The following examples will further illustrate the present invention, but do not limit the present invention thereby.
[0039] The experimental methods in the following examples and comparative examples, unless otherwise specified, are conventional methods in the art. The experimental materials used in the following examples and comparative examples are commercially available unless otherwise specified.
[0040] Chitosan powder was purchased from Sigma Company.
[0041] Tβ4 was purchased from ProSpec Corporation.
[0042] The scanning electron microscope was purchased from Hitachi, Japan, model TM100.
[0043] The preparation method of EDC / NHS cross-linking agent is: dissolve EDC, NHS respectively in 95% ethanol solution to concentration to be 33nmol / L and 8nmol / L, obtain EDC solution and NHS solution, then equal volume of EDC solution and NHS solution mix.
preparation example
[0045] This preparation example is used to illustrate the extraction method of collagen powder.
[0046] (1) Take 10 g of fresh bovine tendon, remove the surrounding connective tissue, and cut the bovine tendon into 1-2mm sized tissue pieces.
[0047] (2) Put the tissue block into 0.1% Na 2 CO 3 Soak in the solution for 2h, rinse with distilled water 3 times, 10min each time.
[0048] (3) Soak the tissue block in 0.5% Tris-HCl (pH=7.5) solution containing 1 mol / L NaCl for 12 hours, rinse with distilled water for 3 times, 10 minutes each time.
[0049] (4) Put the tissue pieces into 250 mL of 0.05 mol / L acetic acid solution containing 500 mg of pepsin, soak at 4°C for 72 hours, and stir repeatedly until fully digested into a transparent paste.
[0050] (5) The mixture was centrifuged at 5000 g for 20 min at 4° C., and the supernatant was taken.
[0051] (6) Perform the operations of steps (4) and (5) on the centrifuged sediment in step (5), and repeat the operation 2 times....
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