Combined Effects of Electrical Stimulation and Protein Coatings on Myotube Formation in a Soft Porous Scaffold

  • Federica IberiteEmail author
  • Irini Gerges
  • Lorenzo Vannozzi
  • Attilio Marino
  • Marco Piazzoni
  • Tommaso Santaniello
  • Cristina Lenardi
  • Leonardo Ricotti
Original Article


Compared to two-dimensional cell cultures, three-dimensional ones potentially allow recreating natural tissue environments with higher accuracy. The three-dimensional approach is being investigated in the field of tissue engineering targeting the reconstruction of various tissues, among which skeletal muscle. Skeletal muscle is an electroactive tissue which strongly relies upon interactions with the extracellular matrix for internal organization and mechanical function. Studying the optimization of myogenesis in vitro implies focusing on appropriate biomimetic stimuli, as biochemical and electrical ones. Here we present a three-dimensional polyurethane-based soft porous scaffold (porosity ~ 86%) with a Young’s modulus in wet conditions close to the one of natural skeletal muscle tissue (~ 9 kPa). To study the effect of external stimuli on muscle cells, we functionalized the scaffold with extracellular matrix components (laminin and fibronectin) and observed an increase in myoblast proliferation over three days. Furthermore, the combination between laminin coating and electrical stimulation resulted in more spread and thicker myotubes compared to non-stimulated samples and samples receiving the single (non-combined) inputs. These results pave the way to the development of mature muscle tissue within three-dimensional soft scaffolds, through the combination of biochemical and electrical stimuli.


Three-dimensional scaffold Biophysical stimulation Skeletal muscle Tissue engineering Polyurethane scaffold 





Dulbecco’s Modified Eagle’s Medium




Extracellular matrix


Electrical stimulation




Fetal bovine serum




Hexamethylene diisocyanate


Interquartile range




Myoblast differentiation medium


Myoblast growth medium


Myoblast seeding medium


Phosphate buffered saline


Polyethylene glycol 6000 g/mol






Scanning electron microscopy


Skeletal muscle


Skeletal muscle tissue engineering





The authors would like to thank Federica Elliot for English language editing and review.

Conflict of interest

The authors declare no competing financial interest.

Supplementary material

10439_2019_2397_MOESM1_ESM.docx (1.6 mb)
Supplementary material 1 (DOCX 1619 kb)


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Copyright information

© Biomedical Engineering Society 2019

Authors and Affiliations

  1. 1.The BioRobotics InstituteScuola Superiore Sant’AnnaPisaItaly
  2. 2.Tensive S.r.lMilanItaly
  3. 3.Smart Bio-InterfacesIstituto Italiano di TecnologiaPontedera, PisaItaly
  4. 4.Interdisciplinary Centre for Nanostructured Materials and Interfaces (CIMaINa) and Department of PhysicsUniversità degli Studi di MilanoMilanItaly
  5. 5.Department of Excellence in Robotics & AIScuola Superiore Sant’AnnaPisaItaly

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