Programming microenvironmental signals with bioactive peptide amphiphiles for skeletal and cardiac myogenesis

Abstract

The extracellular matrix (ECM) is crucial for the coordination and regulation of various cellular processes, including cell adhesion, recruitment, differentiation and death. ECM components structurally support tissue function and regeneration by acting as a substrate for cell migration and differentiation. In addition, by facilitating the fine localization of signals within their structural framework, these components activate receptors on the cell membrane for the initiation of signal transduction cascades. As such, cell-matrix interactions and matrix-associated signals are important for the normal functioning of cells, as well as for natural or artificially assisted tissue regeneration. In keeping with this ECM-centric approach, we designed and synthesized peptide amphiphiles with bioactive epitopes to resemble the native microenvironment of muscle tissue and to examined their potential in the induction of progenitor cell differentiation into skeletal myotubes and cardiac myocytes. The formation of skeletal myotubes was promoted through the use of basal laminamimetic peptide nanofibers inspired by the chemical structures of laminin and fibronectin, two proteins strongly represented in the skeletal muscle extracellular matrix. We demonstrated that our basal lamina mimetic peptide nanofiber system actively interacts with the cells it contains and enhances their differentiation within 3 days. Morphological analysis and immunocytochemical stainings indicated the formation of differentiated myotubes.We also designed glycosaminoglycan-mimetic peptide amphiphiles to mimic the glycosaminoglycans found in the myocardium. Glycosaminoglycans have been reported to play substantial roles in growth factor binding and the induction of angiogenesis, and their mimicry through peptide amphiphile nanofibers is promising as a combined approach for generating multifunctional cardiovascular tissue engineering scaffolds. We demonstrated that peptide nanofibers enhance the adhesion of cells to the surface and induce cardiac myoblast cells to differentiate into cardiomyocytes through both gene expression analysis and immunostainings. In summary, myogenic platforms were developed by programming signal rich environment from self-assembled peptide nanofibers inspired from the components of the ECM to induce the differentiation of cells. These bioactive nanofiber systems serve as promising platforms for muscle tissue engineering applications.