Sphingosine-1-Phosphate Immobilized on Nano-topographical Scaffolds Improve Myogenic Differentiation
The skeletal muscle consists of highly aligned dense cables of collagen fibers with nanometer feature size to support muscle fibers. The skeletal myocyte can be greatly affected to differentiate by their surrounding topographical structure. To improve myogenic differentiation, we fabricated cell culture platform that sphingosine-1-phosphate (S1P) which regulated myocyte behavior is immobilized on a biomimetic nanopatterned polyurethaneacrylate (PUA) substrate using 3,4-dihydroxyphenylalanine (L-DOPA) for providing topographical and biological cues synergistically. In the present study, we hypothesized that cultured C2C12 cells can be induced to synergistically promote myogenic differenntiation on nanopatterned PUA-L-DOPA-S1P. We confirmed that nanopatterned PUA-L-DOPA-S1P has high hydrophilicity with a suitable range of water contact angle and small intensity of phosphate peak (P2p) by analyses of water contact analyzer and X-ray photoelectron spectroscopy. In addition, C2C12 cells culured on nanopatterned PUA-LDOPA- S1P has well-oriented and organized myodubes formed with greater expression of myogenic regulatory factors such as MyoD and MyoG comapred to flat PUA groups. This functional platform which is not only provided topographical and biological cues has a suitable potential function to apply muscle cell niche as similar structure of muscle fiber but also utilized cell behavior within tissue engineered scaffolds and cellular microenvironment.
Keywords 3,4-dihydroxyphenylalanine, myogenic differentiation, myotube formation, nanotopography, sphingosine-1-phosphate