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  • Clinical Research
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Use of arginine–glycine–aspartic acid adhesion peptides coupled with a new collagen scaffold to engineer a myocardium-like tissue graft

Abstract

Background Cardiac tissue engineering might be useful in treatment of diseased myocardium or cardiac malformations. The creation of functional, biocompatible contractile tissues, however, remains challenging. We hypothesized that coupling of arginine–glycine–aspartic acid–serine (RGD+) adhesion peptides would improve cardiomyocyte viability and differentiation and contractile performance of collagen-cell scaffolds.

Methods Clinically approved collagen scaffolds were functionalized with RGD+ cells and seeded with cardiomyocytes. Contractile performance, cardiomyocyte viability and differentiation were analyzed at days 1 and 8 and/or after culture for 1 month.

Results The method used for the RGD+ cell–collagen scaffold coupling enabled the following features: high coupling yields and complete washout of excess reagent and by-products with no need for chromatography; spectroscopic quantification of RGD+ coupling; a spacer arm of 36Å, a length reported as optimal for RGD+-peptide presentation and favorable for integrin-receptor clustering and subsequent activation. Isotonic and isometric mechanical parameters, either spontaneous or electrostimulated, exhibited good performance in RGD+ constructs. Cell number and viability was increased in RGD+ scaffolds, and we saw good organization of cell contractile apparatus with occurrence of cross-striation.

Conclusions We report a novel method of engineering a highly effective collagen-cell scaffold based on RGD+ peptides cross-linked to a clinically approved collagen matrix. The main advantages were cell contractile performance, cardiomyocyte viability and differentiation.

Key Points

  • We propose a new method for tissue engineering to create an efficient and safe contractile tissue by improving the interaction of contractile cells and their surrounding three-dimensional collagen environment

  • The approach increases the functionality of collagen templates by covalent binding and optimal presentation of molecules, such as hormones, growth factors, cytokines and adhesive ligand molecules (i.e. arginine–glycine–aspartic acid peptides)

  • We applied several tools for assessment of myocyte contractility in vitro, in three-dimensional cellular architectures under isometric and isotonic conditions

  • This scaffold might also serve as a platform for improving cellular transplantation in the heart or for engineering a true myocardium equivalent

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Figure 1: The process for coupling adhesive RGD+ peptides to collagen scaffolds.
Figure 2: Contractile performance in RGD+ and control constructs.
Figure 3: Relaxation properties of RGD+ and control constructs.
Figure 4: Comparison of numbers and organization of neonatal rat cardiomyocytes seeded in RGD+ and control scaffolds after 8 days of culture.

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Correspondence to Olivier Schussler.

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Schussler, O., Coirault, C., Louis-Tisserand, M. et al. Use of arginine–glycine–aspartic acid adhesion peptides coupled with a new collagen scaffold to engineer a myocardium-like tissue graft. Nat Rev Cardiol 6, 240–249 (2009). https://doi.org/10.1038/ncpcardio1451

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  • DOI: https://doi.org/10.1038/ncpcardio1451

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