Advertisement

Myoblast adhesion, proliferation and differentiation on human elastin-like polypeptide (HELP) hydrogels

Abstract

Background

The biochemical, mechanical and topographic properties of extracellular matrix are crucially involved in determining skeletal muscle cell morphogenesis, proliferation and differentiation. Human elastin-like polypeptides (HELPs) are recombinant biomimetic proteins designed to mimic some properties of the native matrix protein; when employed as myoblast adhesion substrates, they stimulate in vitro myogenesis. Given the influence that the biophysical properties of extracellular matrix have on skeletal muscle cells, the aim of this work was to investigate the effects of HELP hydrogels on myoblasts’ viability and functions.

Methods

We recently synthesized a novel polypeptide, HELPc, by fusing the elastin-like backbone to a 41aa sequence present in the α2 chain of type IV collagen, containing two arginyl-glycyl-aspartic acid (RGD) motifs. To obtain hydrogels, the enzymatic cross-linking of the HELPc was accomplished by transglutaminase. Here, we employed both non-cross-linked HELPc glass coatings and cross-linked HELPc hydrogels at different monomer densities, as adhesion substrates for C2C12 cells, used as a myoblast model.

Results

By comparing cell adhesion, proliferation and differentiation, we revealed several striking differences. Depending on support rigidity, adhesion to HELPc substrates dictated cell morphology, spreading, focal adhesion formation and cytoskeletal organization. Hydrogels greatly stimulated cell proliferation, particularly in low-serum medium, and partially inhibited myogenic differentiation.

Conclusions

On the whole, the results underline the potential of these genetically engineered polypeptides as a tool for dissecting crucial steps in myogenesis.

J Appl Biomater Funct Mater 2017; 15(1): e43 - e53

Article Type: ORIGINAL RESEARCH ARTICLE

DOI:10.5301/jabfm.5000331

Authors

Paola D’Andrea, Deborah Civita, Michela Cok, Luisa Ulloa Severino, Francesca Vita, Denis Scaini, Loredana Casalis, Paola Lorenzon, Ivan Donati, Antonella Bandiera

Article History

Disclosures

Financial support: This work was supported in part by the European Regional Development Fund (Cross-border cooperation Program Italy-Slovenia 2007-2013, Trans2care strategic project) and by Beneficentia Stiftung (BEN 2014/125), Vaduz, Lichtenstein.
Conflict of interest: The authors declare they have no conflicts of interest.

This article is available as full text PDF.

  • If you are a Subscriber, please log in now.

  • Article price: Eur 36,00
  • You will be granted access to the article for 72 hours and you will be able to download any format (PDF or ePUB). The article will be available in your login area under "My PayPerView". You will need to register a new account (unless you already own an account with this journal), and you will be guided through our online shop. Online purchases are paid by Credit Card through PayPal.
  • If you are not a Subscriber you may:
  • Subscribe to this journal
  • Unlimited access to all our archives, 24 hour a day, every day of the week.

Authors

Affiliations

  • Department of Life Sciences, University of Trieste, Trieste - Italy
  • NanoInnovation Lab at ELETTRA Synchrotron Light Source, Basovizza, Trieste - Italy
  • B.R.A.I.N. Centre for Neuroscience, University of Trieste, Trieste - Italy

Article usage statistics

The blue line displays unique views in the time frame indicated.
The yellow line displays unique downloads.
Views and downloads are counted only once per session.

This article has supplementary materials available to download.

  • If you are a Subscriber, please log in now.

  • Article price: Eur 36,00
  • You will be granted access to the article for 72 hours and you will be able to download any format (PDF or ePUB). The article will be available in your login area under "My PayPerView". You will need to register a new account (unless you already own an account with this journal), and you will be guided through our online shop. Online purchases are paid by Credit Card through PayPal.
  • If you are not a Subscriber you may:
  • Subscribe to this journal
  • Unlimited access to all our archives, 24 hour a day, every day of the week.