title: In vitro myogenesis induced by human recombinant elastin-like proteins.
abstract: Mammalian adult skeletal muscle has a limited ability to regenerate after injury, usage or trauma. A promising strategy for successful regenerative technology is the engineering of bio interfaces that mimic the characteristics of the extracellular matrix. Human elastin-like polypeptides (HELPs) have been synthesized as biomimetic materials that maintain some peculiar properties of the native protein. We developed a novel Human Elastin Like Polypeptide obtained by fusing the elastin-like backbone to a domain present in the α2 chain of type IV collagen, containing two RGD motives. We employed this peptide as adhesion substrate for C2C12 myoblasts and compared its effects to those induced by two other polypeptides of the HELP series. Myoblast adhered to all HELPs coatings, where they assumed morphology and cytoarchitecture that depended on the polypeptide structure. Adhesion to HELPs stimulated at a different extent cell proliferation and differentiation, the expression of Myosin Heavy Chain and the fusion of aligned fibers into multinucleated myotubes. Adhesion substrates significantly altered myotubes stiffness, measured by Atomic Force Microscopy, and differently affected the cells Ca(2+) handling capacity and the maturation of excitation-contraction coupling machinery, evaluated by Ca(2+) imaging. Overall, our findings indicate that the properties of HELP biopolymers can be exploited for dissecting the molecular connections underlying myogenic differentiation and for designing novel substrates for skeletal muscle regeneration.
authors.affiliation: Department of Life Sciences, University of Trieste, I-34127 Trieste, Italy. Electronic address: dandrea@units.it.
authors.forename: Paola
authors.initials: P
authors.lastname: D'Andrea
authors.affiliation: Department of Life Sciences, University of Trieste, I-34127 Trieste, Italy; NanoInnovation Lab at ELETTRA, Synchrotron Light Source S.S. 14 km 163.5, 34012 Basovizza, Trieste, Italy.
authors.forename: Denis
authors.initials: D
authors.lastname: Scaini
authors.affiliation: Department of Life Sciences, University of Trieste, I-34127 Trieste, Italy; NanoInnovation Lab at ELETTRA, Synchrotron Light Source S.S. 14 km 163.5, 34012 Basovizza, Trieste, Italy.
authors.forename: Luisa
authors.initials: L
authors.lastname: Ulloa Severino
authors.affiliation: Department of Life Sciences, University of Trieste, I-34127 Trieste, Italy.
authors.forename: Violetta
authors.initials: V
authors.lastname: Borelli
authors.affiliation: Department of Life Sciences, University of Trieste, I-34127 Trieste, Italy.
authors.forename: Sabina
authors.initials: S
authors.lastname: Passamonti
authors.affiliation: Department of Life Sciences, University of Trieste, I-34127 Trieste, Italy; Centre for Neuroscience B.R.A.I.N., University of Trieste, I-34127 Trieste, Italy.
authors.forename: Paola
authors.initials: P
authors.lastname: Lorenzon
authors.affiliation: Department of Life Sciences, University of Trieste, I-34127 Trieste, Italy.
authors.forename: Antonella
authors.initials: A
authors.lastname: Bandiera
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D002110:Caffeine / Q000494:pharmacology; D002118:Calcium / Q000378:metabolism; D020013:Calcium Signaling / Q000187:drug effects; D002448:Cell Adhesion / Q000187:drug effects; D002454:Cell Differentiation / Q000187:drug effects; D002460:Cell Line; D048430:Cell Shape / Q000187:drug effects; D002470:Cell Survival / Q000187:drug effects; D020099:Coated Materials, Biocompatible / Q000494:pharmacology; D004549:Elastin / Q000737:chemistry* / Q000494:pharmacology; D056966:Excitation Contraction Coupling / Q000187:drug effects; D006801:Humans; D051379:Mice; D018625:Microscopy, Atomic Force; D008969:Molecular Sequence Data; D024510:Muscle Development / Q000187:drug effects*; D018485:Muscle Fibers, Skeletal / Q000166:cytology; D009504:Neutrophils / Q000166:cytology / Q000187:drug effects; D010455:Peptides / Q000138:chemical synthesis / Q000494:pharmacology; D011189:Potassium Chloride / Q000494:pharmacology; D011994:Recombinant Proteins / Q000737:chemistry* / Q000494:pharmacology
chemical_list: D020099:Coated Materials, Biocompatible; D010455:Peptides; D011994:Recombinant Proteins; D002110:Caffeine; D011189:Potassium Chloride; D004549:Elastin; D002118:Calcium
keywords: Biomimetic materials; Cell adhesion; Elastin-like polypeptides; Excitation-contraction coupling; Intracellular calcium; Skeletal muscle regeneration