Background: The topographical cue is major influence on skeletal muscle cell culture because the structure is highly organized and consists of long parallel bundles of multinucleated myotubes that are formed by differentiation and fusion of myoblast satellite cells. In this technical report, we fabricated a multiscale scaffold using electrospinning and poly (ethylene glycol) (PEG) hydrogel micropatterns to monitor the cell behaviors on nano- and micro-alignment combined scaffolds with different combinations of angles. Results: We fabricated multiscale scaffolds that provide biocompatible and extracellular matrix (ECM)-mimetic environments via electrospun nanofiber and PEG hydrogel micro patterning. MTT assays demonstrated an almost four-fold increase in the proliferation rate during the 7 days of cell culture for all of the experimental groups. Cell orientation and elongation were measured to confirm the myogenic potential. On the aligned fibrous scaffolds, more than 90% of the cells were dispersed ± 20° of the fiber orientation. To determine cell elongation, we monitored nuclei aspect ratios. On a random nanofiber, the cells demonstrated an aspect ratio of 1.33, but on perpendicular and parallel nanofibers, the aspect ratio was greater than 2. Myosin heavy chain (MHC) expression was significantly higher i) on parallel compared to random fibers, ii) the 100 μm compared to the 200 μm line pattern. We confirmed the disparate trends of myotube formation that can be provoked through multi-dimensional scaffolds. Conclusion: We studied more favorable environments that induce cell alignment and elongation for myogenesis by combining nano- and micro-scale patterns. The fabricated system can serve as a novel multi-dimensional platform to study in vitro cell behaviors.
Bibliographical noteFunding Information:
This work was supported by the National Research Foundation (NRF) grant funded by the Ministry of Science, ICT and Future Planning (MSIP) (2015R1A2A1A15054532 and 2010 K001430 "Converging Research Center Program") and by a grant (HI15C2807 and HI15C1744) from the Korean Health Technology R&D Project through the Korean Health Industry Development Institute (KHIDI).
© 2017 The Author(s).
All Science Journal Classification (ASJC) codes
- Ceramics and Composites
- Medicine (miscellaneous)
- Biomedical Engineering