To evaluate the cytocompatibil ity of Arg-Gly-Asp-recombinant spider silk protein (pNSR16) / poly vinyl alcohol (PVA) through in vitro cytotoxicity experiment and cell-material co-culture experiment. Methods pNSR16/PVA scaffold and its extraction were prepared by using solvent casting/particulate leaching method, and NIH-3T3 cells were cultivated with the extraction in vitro. The cytotoxicity of scaffold was analyzed using MTT assay 1, 3 and 5 days after culture. Scanning electron microscope and HE staining observation were conducted 2, 4 and 6 days after culturing NIH-3T3 cells on the pNSR16/PVA scaffold. Immunohistochemistry detection was performed 6 days after co-culture. Adhesion, growthand expression of the cells on the scaffold were observed. Results The cytotoxicity of pNSR16/PVA scaffold was in grade 0. Scanning electron microscope observation: the cells covered the surface of the scaffold and were arranged in a directional manner 4 days after co-culture. HE staining: the cells adhered to and grew on the surface of scaffold, and migrated into the scaffold with the increase of culture duration. Immunohistochemistry detection: bFGF was secreted by NIH-3T3 cells, and the cells differentiated normally. Conclusion pNSR16/PVA scaffold has a satisfactory cytocompatibil ity and may be an ideal tissue engineered scaffold materia
Due to its special sequence structure, spider silk protein has unique physical and chemical properties, mechanical properties and excellent biological properties. With the expansion of the application value of spider silk in many fields as a functional material, progress has been made in the studies on the expression of recombinant spider silk proteins through many host systems by gene recombinant techniques. Recombinant spider silk proteins can be processed into high performance fibers, and a wide range of non-fibrous morphologies. Moreover, for their excellent biocompatibility and low immune response they are ideal for biomedical applications. Here we review the process and mechanism of preparation in vitro, chemistry and genetic engineering modification on recombinant spider silk protein.