Manufacturing of Graphene-Nanoplatelet- and Carbon-Nanofiber-Filled PLA Composite Filaments for Tissue Engineering

Electrical stimulation enhances functionality and accelerates maturation in biofabricated tissues, which are particularly important for muscle tissue engineering applications. Accordingly, there is demand for 3D-printable electrically conductive cytocompatible scaffolds that enable patient-specific geometries and localized electrical stimulation, as well as incorporate further maturation-promoting geometrical cues. Filament-based scaffolds from fused filament fabrication could overcome current limitations in geometric freedom, size and partially cytotoxic additives. In this study, biodegradable polylactic acid (PLA)-based conductive filaments incorporating graphene nanoplatelets (GNPs) or carbon nanofibers (CNFs) were developed via melt-mixing extrusion to possibly enable the electrical functionalization of muscle scaffolds. A two-stage process combining twin-screw and single-screw extrusion was preferred to allow for higher filler incorporation. Filament morphology, printability, electrical conductivity, and cytocompatibility were systematically evaluated. Homogeneous filaments containing up to 16 wt.% GNPs or 3.6 wt.% CNFs were successfully produced and processed by fused filament fabrication into scaffold geometries supporting myoblast orientation. Electrical conductivity was measured above 16 wt.% GNPs, with up to 2.7 µS/m, with printed constructs capable of connecting a circuit. GNP-based filaments were cytocompatible, supporting myoblast attachment and elongated morphology. An adjustable electrical stimulation setup demonstrated improved muscle maturation and contractile responses of C2C12 myoblasts, highlighting biodegradable conductive filaments' potential for electrically active muscle tissue scaffolds.