Silk fibroin/collagen scaffold material loaded with growth factors and preparation method
A collagen and growth factor technology, applied in medical science, prostheses, etc., can solve the problems of not reaching the effective concentration, losing biological activity, etc., to solve the problem of short half-life in the body, prolonging the functional effect, prolonging the full Play the effect
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Embodiment 1
[0025] Example 1: Preparation of loose layer silk fibroin / collagen film and dense layer silk fibroin / collagen film
[0026] 1. Preparation of silk fibroin
[0027] Cut silkworm cocoons into pieces and add 0.5% Na 2 CO 3 solution, in a constant temperature water bath at 98±2°C for 30 min, twice in total. After taking it out, wash it twice with boiling water, and then wash it 8 times with deionized water. Dry in an oven at 60° C. to obtain dry silk fibroin (ie, silk fibroin). Weigh the silk fibroin silk after silk degumming, use CaCl 2 / H 2 O / C 2 h 5 OH solution (molar ratio 1:8:2) was dissolved in a constant temperature water bath at (78±2)°C, and the bath ratio was 1:25. Filter with gauze to obtain the silk fibroin stock solution. Dialyze with a fibrous semi-permeable membrane dialysis bag with a cut-off molecular weight of 8000D-14000D for 3 days to remove calcium chloride and ethanol. Filter to obtain silk fibroin solution.
[0028] 2. Collagen preparation
[002...
Embodiment 2
[0035] Example 2: Preparation of TGF-β1 Nanoparticles Encapsulated
[0036] Polylysine is dissolved in deionized water with pH=7.4, the content is 1 mg.ml -1 . Dissolve TGF-β1 in 0.2 ML water, add 5 mL, 1 mg.ml -1 The sodium heparin solution was mixed overnight. Add TGF-β1-heparin sodium solution to 2 mL, 1 mg.ml -1 In the PLL solution, shake on a vortex mixer for 15 s, and then incubate at room temperature for 30 min. TGF-β1-PLL-heparin sodium nanoparticles can be obtained.
Embodiment 3
[0037] Example 3: Preparation of silk fibroin / collagen scaffold material loaded with TGF-β1
[0038] The cross-linked loose layer silk fibroin / collagen membrane prepared in Example 1 was wetted with double-distilled water and spread flat, and the PLL-heparin nanoparticle solution loaded with TGF-β1 prepared in Example 2 was added dropwise on the surface, 4- Air-dry at 8°C until there is no flowing liquid on the surface, then wet the cross-linked dense layer silk fibroin / collagen membrane and spread it on the upper surface of the cross-linked loose layer silk fibroin / collagen membrane, and air-dry at 4-8°C to obtain TGF-β1 silk fibroin / collagen scaffold material.
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