A kind of preparation method of the functional silk fibroin membrane that is beneficial to cell adhesion

Abstract

The invention discloses a preparation method of a functional silk fibroin film which is beneficial to cell adhesion. The steps include: transfecting Escherichia coli with a prokaryotic system expression vector carrying (‑RGD‑)4 polypeptide or (‑RGD‑)8 polypeptide gene Cells, induced and cultured for several hours; centrifuge the cell culture liquid, collect and crush the bacterial cells, purify the crushed protein supernatant, collect by enzymatic digestion in one step, and freeze-dry to obtain (‑RGD‑)4 polypeptide powder or (‑RGD ‑)8 Polypeptide powder; degumming, dissolving, dialysis, filtering and concentrating silkworm silk or silkworm cocoons, pouring them into a container with a certain area and air-drying to obtain silkworm fibroin film; immersing the silkworm silk film in excess cross-linking agent solution Then add (‑RGD‑)4 polypeptide or (‑RGD‑)8 polypeptide and react overnight at 4°C to obtain a functional silk fibroin film. The functional silk fibroin membrane prepared by the invention has excellent cell adhesion performance, belongs to biomedical materials, has no cytotoxicity, has recognition sites of cell adhesion proteins, and is beneficial to cell adhesion, growth, and promotion of defect tissue healing.

Description

technical field

[0001] The invention relates to a preparation method of a silk fibroin material applied in the field of biomedicine, in particular to a preparation method of a functional silk fibroin film favorable for cell adhesion. Background technique

[0002] The RGD sequence is composed of arginine, glycine and aspartic acid, and widely exists in a variety of extracellular matrices, such as collagen, fibronectin, laminin, vitronectin, elastin, etc. A variety of integrins specifically bind, contribute to cell adhesion, and promote the adhesion and growth of adherent cells. Silk membranes chemically modified by RGD and other modified materials such as polymers can improve the adhesion, diffusion and proliferation of cells on materials (such as J Biomed MaterRes A, 2003, 67 (2), 559-570; Biomaterials, 2002, 23: 4315-4323, etc.). RGD peptide can also competitively inhibit the binding of various adhesion proteins such as fibrin to platelets and the adhesion of platelets to...

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