Multifunctional scaffold based on Ti3C2Tx/PCL nanofibers composite hydrogel directs myogenic differentiation and microenvironment remodeling for volumetric muscle loss

Volumetric muscle loss (VML) regeneration remains clinically challenging, primarily due to compromised myogenic differentiation and an oxidative-inflammatory microenvironment that impedes regeneration. Consequently, timely resolution of inflammation and dynamic remodeling of the immune niche are imperative for functional VML recovery. Ti 3 C 2 T x MXene, a two-dimensional nanomaterial, demonstrates significant potential in biomedical applications owing to its anti-inflammatory and immunomodulatory properties. However, oxidative degradation and hydrolysis occur upon aqueous exposure, leading to compromised bioactivity. To preserve structural integrity and mitigate oxidative degradation of Ti 3 C 2 T x MXene, it was encapsulated in electrospun fibers to fabricate Ti 3 C 2 T x MXene/PCL membrane (MP). This biomimetic multifunctional scaffold was fabricated via hydrogen bond-driven self-assembly, integrating alternating layers of MP membranes and hyaluronic acid-catechol/4-arm-PEG (HP) hydrogel, and is hereinafter referred to as the MPHP scaffold. In vitro analyses demonstrated enhanced cellular adhesion and myogenic differentiation (evidenced by > 3.9-fold upregulation of myogenic differentiation (MyoD) and >2.9-fold upregulation of myogenin (MyoG)), confirming potent myogenesis-promoting capabilities. In vitro and in vivo analyses consistently demonstrated the scaffold's capacity to reprogram macrophage polarization to regenerative M2 phenotypes, effectively remodeling the regenerative niche. During the remodeling phase, it accelerated myotube fusion and maturation, while concurrently promoting angiogenesis and neuromuscular junction reinnervation. These coordinated mechanisms restored structural and functional muscle integrity, with treated VML models exhibiting a 38% increase in exercise tolerance versus controls. The MPHP scaffold thus represents a multifunctional biomimetic platform for VML regeneration, demonstrating significant translational potential in tissue engineering. Moreover, this encapsulation strategy preserves Ti 3 C 2 T x MXene bioactivity and expands its therapeutic applicability beyond conventional limitations.