3D Rotary Wet-Spinning (RoWS) Biofabrication Directly Affects Proteomic Signature and Myogenic Maturation in Muscle Pericyte–Derived Human Myo-Substitute

Abstract

Skeletal muscle tissue engineering (SMTE) has recently emerged to address major clinical challenges such as volumetric muscle loss (VML). Here, we report a rotary wet-spinning (RoWS) biofabrication technique for producing human myo-substitutes with biomimetic architectures and functions. Here, we demonstrate how the proposed technique may be used to establish a well-tailored, anisotropic microenvironment that promotes myogenic differentiation of human skeletal muscle–derived pericytes (hPeri). Using high-resolution mass spectrometry–based proteomics with the integration of literature-derived signaling networks, we uncovered that (i) a 3D biomimetic matrix environment (PEG-fibrinogen) confers a less mitogenic microenvironment compared to standard 2D cultures, favoring the formation of contractile-competent bundles of pericyte-derived myotubes in an anchoring-independent 3D state and (ii) the RoWS method promotes an upregulation of muscle matrix structural protein besides increasing contractile machinery proteins with respect to 3D bulk cultures. Finally, in vivo investigations demonstrate that the 3D-biofabricated myo-substitute is fully compatible with the host ablated muscular tissue, exhibiting myo-substitute engraftment and muscle regeneration in a mouse model of VML. Overall, the results show that RoWS offers a superior capability for controlling the myogenic differentiation process on a macroscale and, with future refining, may have the potential to be translated into clinical practice.