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Ann Biomed Eng. 2016 Jun;44(6):2036-48. doi: 10.1007/s10439-016-1586-6. Epub 2016 Mar 16.

Cell-Instructive Graphene-Containing Nanocomposites Induce Multinucleated Myotube Formation.

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Department of Pharmaceutical Sciences, School of Pharmacy, University of Pittsburgh, Pittsburgh, PA, 15261, USA.
Center for Biologic Imaging, University of Pittsburgh School of Medicine, Pittsburgh, PA, 15261, USA.
Departments of Cell Biology and Pathology, University of Pittsburgh, Pittsburgh, PA, 15261, USA.
Department of Pharmaceutical Sciences, School of Pharmacy, University of Pittsburgh, Pittsburgh, PA, 15261, USA.
Department of Bioengineering, University of Pittsburgh, Pittsburgh, PA, 15219, USA.
McGowan Institute for Regenerative Medicine, Pittsburgh, PA, 15260, USA.


Myoblast differentiation is a key step in myogenesis and has long been considered to be controlled mainly by biochemical cues such as growth factors. However, the tissue engineering approaches based on biochemical cues demonstrate low reproducibility as a precise spatial control over their bioactivity is challenging. Recently, substrate micro/nano-structure and electro-responsive properties are recognized for their important roles in myoblast differentiation. In this study, we hypothesized that engineering biophysical features such as nano/micro-fibrous structure and conductive properties into a single biomaterial scaffold will instruct the myoblasts to differentiate into multinucleated myotubes even in the absence of differentiation media. We fabricated nanocomposite scaffolds composed of conductive graphene nanosheets and polycaprolactone (PCL), a widely used biocompatible material. The resulting graphene-PCL scaffolds possess excellent conductivity due to graphene nanosheets and great processability, biodegradability and elastic mechanical properties conferred by PCL. Additionally, physicochemical and mechanical properties of nanocomposite scaffolds can be tuned by varying graphene concentration. Further, graphene-PCL nanocomposites and their 8-week degradation products exhibited remarkable cytocompatibility and promoted adhesion and proliferation of C2C12 mouse myoblast cells. Importantly, these nanocomposite scaffolds induced graphene concentration-dependent differentiation of C2C12 cells into multinucleated myotubes even in normal growth media suggesting their cell-instructive potential. Thus, graphene-PCL nanocomposite scaffolds can serve as a strategy to promote skeletal muscle regeneration without biochemical cues.


Biologic-free; Cell-instructive scaffolds; Electrospinning; Graphene nanosheet; Myoblast differentiation; Polycaprolactone

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