Objective To explore the construction and biocompatibility in vitro evaluation of the electrospun-graphene (Gr)/silk fibroin (SF) nanofilms. Methods The electrostatic spinning solution was prepared by dissolving SF and different mass ratio (0, 5%, 10%, 15%, and 20%) of Gr in formic acid solution. The hydrophilia and hydrophobic was analyzed by testing the static contact angle of electrostatic spinning solution of different mass ratio of Gr. Gr-SF nanofilms with different mass ratio (0, 5%, 10%, 15%, and 20%, as groups A, B, C, D, and E, respectively) were constructed by electrospinning technology. The structure of nanofilms were observed by optical microscope and scanning electron microscope; electrochemical performance of nanofilms were detected by cyclic voltammetry at electrochemical workstation; the porosity of nanofilms were measured by n-hexane substitution method, and the permeability were observed; L929 cells were used to evaluate the cytotoxicity of nanofilms in vitro at 1, 4, and 7 days after culture. The primary Sprague Dawley rats’ Schwann cells were co-cultured with different Gr-SF nanofilms of 5 groups for 3 days, the morphology and distribution of Schwann cells were identified by toluidine blue staining, the cell adhesion of Schwann cells were determined by cell counting kit 8 (CCK-8) method, the proliferation of Schwann cells were detected by EdU/Hoechst33342 staining. Results The static contact angle measurement confirmed that the hydrophilia of Gr-SF electrospinning solution was decreased by increasing the mass ratio of Gr. Light microscope and scanning electron microscopy showed that Gr-SF nanofilms had nanofiber structure, Gr particles could be dispersed uniformly in the membrane, and the increasing of mass ratio of Gr could lead to the aggregation of particles. The porosity measurement showed that the Gr-SF nanofilms had high porosity (>65%). With the increasing of mass ratio of Gr, the porosity and conductivity of Gr-SF nanofilm increased gradually, the value in the group A was significantly lower than those in groups C, D, and E (P<0.05). In vitro L929 cells cytotoxicity test showed that all the Gr-SF nanofilms had good biocompatibility. Toluidine blue staining, CCK-8 assay, and EdU/Hoechst33342 staining showed that Gr-SF nanofilms with mass ratio of Gr less than 10% could support the survival and proliferation of co-cultured Schwann cells. Conclusion The Gr-SF nanofilm with mass ratio of Gr less than 10% have proper hydrophilia, conductivity, porosity, and other physical and chemical properties, and have good biocompatibility in vitro. They can be used in tissue engineered nerve preparation.
ObjectiveTo explore the preparation method, physical and chemical properties, and biocompatibility of a conductive composite scaffold based on polypyrrole/silk fibroin (PPy/SF) fiber with " shell-core” structure, and to provide a preliminary research basis for the application in the field of tissue engineered neuroscience.Methods The conductive fibers with " shell-core” structure were prepared by three-dimensional printing combined with in-situ polymerization. PPy/SF fiber-based conductive composite scaffolds were formed by electrospinning. In addition, core-free PPy conductive fibers and SF electrospinning fibers were prepared. The stability, biomechanics, electrical conductivity, degradation performance, and biological activity of each material were tested to analyze the comprehensive properties of fiber-based conductive composite scaffolds.ResultsCompared with pure core-free PPy conductive fibers and SF electrospinning fibers, the PPy/SF fiber-based conductive composite scaffolds with " shell-core” structure could better maintain the stability performance, enhance the mechanical stretchability of the composite scaffolds, maintain long-term electrical activity, and improve the anti-degradation performance. At the same time, PPy/SF conductive composite scaffolds were suitable for NIH3T3 cells attachment, conducive to cell proliferation, and had good biological activity.ConclusionPPy/SF fiber-based conductive composite scaffolds meet the needs of conductivity, stability, and biological activity of artificial nerve grafts, and provide a new idea for the development of a new generation of high-performance and multi-functional composite materials.