An electro-responsive functional neurovascularized engineered muscle tissue

Integrate multiple advanced technologies to rebuild a functional tissue with both neurotization and vascularization in the field of tissue engineering. Based on light-induced anisotropic cell sheets (ACS) technology, which enabled the alignment of C2C12 myoblasts on the surface of titanium dioxide nanodots. Then PC12 neural cells were added to co-culture with C2C12s, and the optimal ratio was selected to construct an aligned neuralized muscle structure. Meanwhile, endothelial cells were induced to form the aligned vascular network. High-precision 3d printing technology was used to construct the flexible electrodes, and the above structures were assembled by hydrogel to finally form the composite constructs. The constructs were implanted in nude mice volumetric muscle loss (VML) models for validation, and in vivo electrical stimulation (ES) was performed during recovery. After 4 weeks, gait analysis techniques were used to assess the efficacy of functional recovery according to gait orderliness, print area and intensity. Muscle samples were collected for histology (H&E, MTS) and immunofluorescence staining (MHC, βIIIT, CD31) to evaluate muscle, nerve, and vascular regeneration. The engineered muscle implanted group showed better muscle, nerve and vascular regeneration ability, and the assistance of electrical stimulation further enhanced muscle functional restoration. The comprehensive repair strategy of electro-responsive functional neurovascularized engineered muscle (EFNEM) combined with in vivo ES can achieve tissue and functional recovery after VML, providing new insights into the field of tissue reconstruction.