Abstract
The degeneration or loss of skeletal muscles, which can be caused by traumatic injury or disease, impacts most aspects of human activity. Among various techniques reported to regenerate skeletal muscle tissue, controlling the external cellular environment has been proven effective in guiding muscle differentiation. In this study, we report a nano-sized graphene oxide (sGO)-modified nanopillars on microgroove hybrid polymer array (NMPA) that effectively controls skeletal muscle cell differentiation. sGO-coated NMPA (sG-NMPA) were first fabricated by sequential laser interference lithography and microcontact printing methods. To compensate for the low adhesion property of polydimethylsiloxane (PDMS) used in this study, graphene oxide (GO), a proven cytophilic nanomaterial, was further modified. Among various sizes of GO, sGO (< 10 nm) was found to be the most effective not only for coating the surface of the NM structure but also for enhancing the cell adhesion and spreading on the fabricated substrates. Remarkably, owing to the micro-sized line patterns that guide cellular morphology to an elongated shape and because of the presence of sGO-modified nanostructures, mouse myoblast cells (C2C12) were efficiently differentiated into skeletal muscle cells on the hybrid patterns, based on the myosin heavy chain expression levels. Therefore, the developed sGO coated polymeric hybrid pattern arrays can serve as a potential platform for rapid and highly efficient in vitro muscle cell generation.
Highlights
Skeletal muscle is one of the major components in the human body that controls most of the body motions and movements [1, 2]
We report a new platform consisting of nanopillar arrays, nano-sized graphene oxide, and microgrooves for highly efficient skeletal muscle cell differentiation (Fig. 1)
The micropattern, nanopattern, and graphene oxide (GO) were employed in the hybrid pattern array for guiding cell differentiation and function
Summary
Skeletal muscle is one of the major components in the human body that controls most of the body motions and movements [1, 2]. We report a new platform consisting of nanopillar arrays, nano-sized graphene oxide (sGO), and microgrooves for highly efficient skeletal muscle cell differentiation (Fig. 1). The laser interference lithography technique is first applied to the photoresist (PR)-coated silicon micropattern mold to generate periodic homogeneous nanohole patterns. The polydimethylsiloxane (PDMS) polymer is applied to the nanohole-modified silicon mold to reversely replicate the structure, resulting in the generation of nanopillars on microgroove hybrid polymer array (NMPA). Graphene oxide (GO) of different sizes are applied and coated to the substrates to enhance the cell adhesion on the micropatterned polymeric arrays along with the nanopatterns [33]. The efficiency of skeletal muscle cell differentiation of C2C12 on differently fabricated platforms is evaluated based on the immunofluorescence images using myosin heavy chain (MHC) as a myogenesis marker. The topography of the fabricated nanopillars on microgroove was observed by atomic force microscopy (AFM)
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