1. In vitro myogenesis induced by human recombinant elastin-like proteins
- Author
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Antonella Bandiera, Sabina Passamonti, Denis Scaini, Luisa Ulloa Severino, Paola Lorenzon, Paola D'Andrea, Violetta Borelli, D'Andrea, Paola, Scaini, Deni, ULLOA SEVERINO, Luisa, Borelli, Violetta, Passamonti, Sabina, Lorenzon, Paola, and Bandiera, Antonella
- Subjects
Neutrophils ,Biomimetic materials ,Muscle Fibers, Skeletal ,Microscopy, Atomic Force ,Muscle Development ,Potassium Chloride ,Extracellular matrix ,Mice ,Coated Materials, Biocompatible ,Skeletal muscle regeneration ,Myosin ,Myocyte ,Cell adhesion ,Elastin-like polypeptides ,Excitation-contraction coupling ,Intracellular calcium ,Excitation Contraction Coupling ,biology ,Myogenesis ,Cell Differentiation ,Recombinant Proteins ,Cell biology ,medicine.anatomical_structure ,Biochemistry ,Mechanics of Materials ,C2C12 ,Materials science ,Cell Survival ,Molecular Sequence Data ,Biophysics ,Bioengineering ,Cell Line ,Biomaterials ,Caffeine ,medicine ,Cell Adhesion ,Animals ,Humans ,Amino Acid Sequence ,Calcium Signaling ,Cell Shape ,Elastin-like polypeptides Biomimetic materials Cell adhesion Skeletal muscle regeneration Excitation-contraction coupling Intracellular calcium ,Skeletal muscle ,Elastin ,Ceramics and Composites ,biology.protein ,Calcium ,Peptides - 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.
- Published
- 2015