Fabrication of skeletal muscle constructs by topographic activation of cell alignment

Abstract

Skeletal muscle fiber construction for tissue-engineered grafts requires assembly of unidirectionally aligned juxtaposed myotubes. To construct such a tissue, a polymer microchip with linearly aligned microgrooves was fabricated that could direct myoblast adaptation under stringent conditions. The closely spaced microgrooves fabricated by a modified replica molding process guided linear cellular alignment. Examination of the myoblasts by immunofluorescence microscopy demonstrated that the microgrooves with subcellular widths and appropriate height-to-width ratios were required for practically complete linear alignment of myoblasts. The topology-dependent cell alignment encouraged differentiation of myoblasts into multinucleate, myosin heavy chain positive myotubes. The monolayer of myotubes formed on the microstructured chips allowed attachment, growth and differentiation of subsequent layers of linearly arranged myoblasts, parallel to the primary monolayer of myotubes. The consequent deposition of additional myoblasts on the previous layer of myotubes resulted in three-dimensional multi-layered structures of myotubes, typical of differentiated skeletal muscle tissue. The findings demonstrate that the on-chip device holds promise for providing an efficient means for guided muscle tissue construction.

Analysis of high-throughput screening reveals the effect of surface topographies on cellular morphology

“…muscle [42] and blood vessel regeneration [43]. Cell guidance through surface topographies has been researched using groove topographies (e.g.…”

Anisotropically oriented electrospun matrices with an imprinted periodic micropattern: a new scaffold for engineered muscle constructs

“…Furthermore, Zhao et al (2009) initiated a double seeding strategy, where additional myoblasts were seeded on differentiated myotubes cultured on micropatterned PDMS substrates. The additional layer of myotubes differentiated in a much quicker time-frame and resulted in an increased myotube width in comparison to the first layer.…”

Bioinspired Nanocomposite Hydrogels with Highly Ordered Structures

“…[13,26] From molecular assemblies to highly aligned extracellular fibrils and cells, a multilevel ordered hierarchy in these biological soft tissues is crucial for their biological function. [13,26,27] For example, skeletal muscles act as core actuation systems; in these muscles, collagen self-assembles into 1D fiber bundles, and these bundles gather directionally. [13,27] Such ordered structures are essential to generate unique anisotropic functions, such as directional motion and shock absorbance.…”

“…[13,26,27] For example, skeletal muscles act as core actuation systems; in these muscles, collagen self-assembles into 1D fiber bundles, and these bundles gather directionally. [13,27] Such ordered structures are essential to generate unique anisotropic functions, such as directional motion and shock absorbance. Corneas with laminated alignment structures are one of the important components in eyes www.advmat.de www.advancedsciencenews.com and can exhibit high mechanical performance.…”