title: Silk fibroin scaffolds with muscle-like elasticity support in vitro differentiation of human skeletal muscle cells.
abstract: Human adult skeletal muscle has a limited ability to regenerate after injury and therapeutic options for volumetric muscle loss are few. Technologies to enhance regeneration of tissues generally rely upon bioscaffolds to mimic aspects of the tissue extracellular matrix (ECM). In the present study, silk fibroins from four Lepidoptera (silkworm) species engineered into three-dimensional scaffolds were examined for their ability to support the differentiation of primary human skeletal muscle myoblasts. Human skeletal muscle myoblasts (HSMMs) adhered, spread and deposited extensive ECM on all the scaffolds, but immunofluorescence and quantitative polymerase chain reaction analysis of gene expression revealed that myotube formation occurred differently on the various scaffolds. Bombyx mori fibroin scaffolds supported formation of long, well-aligned myotubes, whereas on Antheraea mylitta fibroin scaffolds the myotubes were thicker and shorter. Myotubes were oriented in two perpendicular layers on Antheraea assamensis scaffolds, and scaffolds of Philosamia/Samia ricini (S. ricini) fibroin poorly supported myotube formation. These differences were not caused by fibroin composition per se, as HSMMs adhered to, proliferated on and formed striated myotubes on all four fibroins presented as two-dimensional fibroin films. The Young's modulus of A. mylitta and B. mori scaffolds mimicked that of normal skeletal muscle, but A. assamensis and S. ricini scaffolds were more flexible. The present study demonstrates that although myoblasts deposit matrix onto fibroin scaffolds and create a permissive environment for cell proliferation, a scaffold elasticity resembling that of normal muscle is required for optimal myotube length, alignment, and maturation. © 2016 The Authors Journal of Tissue Engineering and Regenerative Medicine Published by John Wiley & Sons Ltd. StartCopTextStartCopText© 2016 The Authors Journal of Tissue Engineering and Regenerative Medicine Published by John Wiley & Sons Ltd.
authors.affiliation: School of Biomedical Science, CHIRI Biosciences Research Precinct, Faculty of Health Sciences, Curtin University, Perth, Western Australia.
authors.forename: Vishal
authors.initials: V
authors.lastname: Chaturvedi
authors.affiliation: Department of Biotechnology, Indian Institute of Technology, Kharagpur, West Bengal, India.
authors.forename: Deboki
authors.initials: D
authors.lastname: Naskar
authors.affiliation: School of Biomedical Science, CHIRI Biosciences Research Precinct, Faculty of Health Sciences, Curtin University, Perth, Western Australia.
authors.forename: Beverley F
authors.initials: BF
authors.lastname: Kinnear
authors.affiliation: Nanochemistry Research Institute, Faculty of Science, Engineering and Computing, Curtin University, Perth, Western Australia.
authors.forename: Elizabeth
authors.initials: E
authors.lastname: Grenik
authors.affiliation: School of Biomedical Science, CHIRI Biosciences Research Precinct, Faculty of Health Sciences, Curtin University, Perth, Western Australia.
authors.forename: Danielle E
authors.initials: DE
authors.lastname: Dye
authors.affiliation: School of Anatomy, Physiology and Human Biology, University of Western Australia, Perth, Western Australia.
authors.forename: Miranda D
authors.initials: MD
authors.lastname: Grounds
authors.affiliation: Department of Biotechnology, Indian Institute of Technology, Kharagpur, West Bengal, India.
authors.forename: Subhas C
authors.initials: SC
authors.lastname: Kundu
authors.affiliation: School of Biomedical Science, CHIRI Biosciences Research Precinct, Faculty of Health Sciences, Curtin University, Perth, Western Australia.
authors.forename: Deirdre R
authors.initials: DR
authors.lastname: Coombe
mesh_terms: D002454:Cell Differentiation; D002478:Cells, Cultured; D004548:Elasticity; D005109:Extracellular Matrix / Q000737:chemistry*; D005349:Fibroins / Q000737:chemistry*; D006801:Humans; D018482:Muscle, Skeletal / Q000166:cytology / Q000378:metabolism*; D032448:Myoblasts, Skeletal / Q000166:cytology / Q000378:metabolism*; D054457:Tissue Scaffolds / Q000737:chemistry*
chemical_list: D005349:Fibroins
keywords: biomaterials; elasticity; extracellular matrix; myotubes; primary human myoblasts; silk fibroin; skeletal muscle tissue engineering