Electrospun nanohybrid hydrogels for enhanced differentiation of myoblasts

Abstract

Enhancement of myogenic differentiation was accomplished with nanohybrid hydrogels composed of fragmented electrospun nanofibrils (NFs) and gelatin/alginate hydrogels. Electrospun nanofibrous meshes were fragmented and hydrolyzed to short fragments of NFs and subsequently embedded in gelatin/alginate hydrogels and further crosslinked with C2C12 cells. Cell-incorporated NF-embedded hydrogels exhibited a lower swelling ratio and higher mechanical properties as compared to hydrogels without NF due to the rigidity of NF and increased matrix-to-cell interaction. Viability of C2C12 cells in the hydrogels gradually increased in proportion to the amount of NF in the hydrogels. Both confocal microscopy and myogenic gene expression analysis indicated that the formation and maturation of myotubes and the expression of muscle motor proteins were significantly accelerated when NF-embedded hydrogels were employed. Contraction behavior of matured myotubes under electrical stimulation revealed that myotubes in NF-embedded hydrogels exhibited a longer contraction length than that of myotubes in hydrogels without NF. Thus, the nanohybrid matrix demonstrates the potential of nanofiber-embedded hydrogels for engineering myotube formation with topographical cues.