Chitosan-sericin composite biological scaffold as well as preparation method and application thereof
A technology of sericin and bio-scaffold, which is applied in the field of medical biocomposite materials, can solve the problems of wasting natural resources, achieve good biocompatibility, and prevent tissue sticking
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Embodiment 1
[0068] Example 1 Preparation of chitosan-sericin composite bioscaffold
[0069] 1. Selection of silkworm cocoons:
[0070] The silkworm cocoons of silkworm silk fibroin-deficient mutant varieties were selected (purchased from the Sericulture Research Institute of the Chinese Academy of Agricultural Sciences, and the silkworm silk fibroin-deficient mutant varieties were preserved in the National Silkworm Resource Preservation Center of the Sericulture Research Institute of the Chinese Academy of Agricultural Sciences, and the preservation number (or product number) 185Nd-s, 140Nd-s, 139Nd-s As the raw material, the main chemical composition is: sericin.
[0071] 2. Choice of chitosan:
[0072] Chitosan was purchased from Sigma with a purity greater than 75%.
[0073] 3. Extraction and separation of sericin
[0074] 1) Weigh 1 g of silkworm cocoons of silk fibroin-deficient mutant varieties and cut them into 1 mm 2 Left and right fragments were placed in a clean beaker. Th...
Embodiment 2
[0090] Example 2 Performance analysis of chitosan-sericin composite bioscaffold
[0091] The chitosan-sericin composite bioscaffold prepared in embodiment 1 is carried out following performance analysis, specifically:
[0092] 1. Macro structure
[0093] Using the selected mold, the resulting chitosan-sericin composite scaffold was prepared into a sheet-like structure with a diameter of 1 cm and a height of 2-3 mm, such as figure 2 shown.
[0094] 2. Microstructure.
[0095] The chitosan-sericin composite bioscaffold was observed under a scanning electron microscope (ULTRA PLUS-43-13, Zeiss, Germany). Such as image 3 As shown, the SEM images of the cross-sections of five chitosan-sericin composite bioscaffolds with different ratios, the scale is 500 microns, and the interior of the scaffold is a multi-void structure, CS100 / SS0, CS80 / SS20, CS50 / SS50, CS20 / SS80 The average pore diameters of the five composite scaffolds, CS0 / SS100, are 157.69 μm, 130.55 μm, 146.27 μm, 136....
Embodiment 3
[0109]Example 3 Evaluation of Composite Biological Scaffold Supporting Nerve Cell Survival and Proliferation Ability
[0110] Use the aseptic chitosan solution prepared in Example 1 and the sericin aqueous solution to directly add in the cell well plate according to CS100 / SS0, CS80 / SS20, CS50 / SS50, CS20 / SS80, CS0 / SS100 five kinds of ratios, each Add genipin aqueous solution in various proportions, the amount added is 330 uL of genipin aqueous solution with a concentration of 0.01 g / mL per 1 mL of mixed solution, at 37 o C for 8 hours, followed by freezing at -80°C for 4 hours, and then freeze-drying in a vacuum freeze dryer to obtain a freeze-dried chitosan-sericin composite bioscaffold; Mouse Schwann cells (RSC96) were suspended, blown up, and seeded on cell culture plates covered with freeze-dried scaffolds. The medium used for cell culture was DMEM (Dulbecco's Modified Eagle's Medium) medium, 5% fetal bovine serum was added to the medium, and the cells were placed in a cel...
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