Skeletal myogenesis on highly orientated microfibrous polyesterurethane scaffolds

Abstract

Skeletal myogenesis is a complex process, which is known to be intimately depending on an optimal outside-in substrate-cell signaling. Current attempts to reproduce skeletal muscle tissue in vitro using traditional scaffolds mainly suffer from poor directionality of the myofibers, resulting in an ineffective vectorial power generation. In this study, we aimed at investigating skeletal myogenesis on novel biodegradable microfibrous scaffolds made of DegraPol, a block polyesterurethane previously demonstrated to be suitable for this application. DegraPol was processed by electrospinning in the form of highly orientated ("O") and nonorientated ("N/O") microfibrous meshes and by solvent-casting in the form of nonporous films ("F"). The effect of the fiber orientation at the scaffold surface was evaluated by investigating C2C12 and L6 proliferation (via SEM analysis and alamarBlue test) and differentiation (via RT-PCR analysis and MHC immunostaining). We demonstrated that highly orientated elastomeric microfibrous DegraPol scaffolds enable skeletal myogenesis in vitro by aiding in (a) myoblast adhesion, (b) myotube alignment, and (c) noncoplanar arrangement of cells, by providing the necessary directional cues along with architectural and mechanical support.

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

“…In addition, sub-micrometer topographies displaying groove sizes of 450-900 nm were found to promote myoblast alignment and fusion into longer, more regularly shaped myotubes in comparison with flat substrates. Similar to reports of substrates with grooves of various widths and depths, electrospun fibres of various polymers in the nanometre or micrometre range were reported to induce spatial myoblast orientation and fusion into multinucleated myotubes (Choi et al 2008, Riboldi et al 2008, Aviss et al 2010. Various studies led to the hypothesis that a wide range of fibre diameter (300 nm to 12 μm) can induce myoblast orientation.…”

Microfeature guided skeletal muscle tissue engineering for highly organized 3-dimensional free-standing constructs

“…We hypothesized that aligning myoblasts and myotubes at the microscale prior to macroscopic construct formation would better mimic the physiologic process of muscle formation and enhance structural organization and force production. Current efforts for aligning cells for use in tissue engineering mostly encompass scaffolds and micropatterned surfaces [3][4][5][6][7]. The issue with using such synthetic materials is that they are undesired in the final tissue, and are difficult or impossible to remove.…”

Ascorbic Acid in Polyurethane Systems for Tissue Engineering

“…In this respect, the range of mechanical and morphological properties that can be obtained with PUR is significantly larger than with commonly used medical grade biodegradable polymers for example: PLA, PGA, poly(DL-lactide) (PDLLA) [39][40][41][42]. Many applications of segmented PUR in TE field, such as cardiovascular TE [34,43,44] musculoskeletal applications (anterior cruciate ligament) [42], knee joint meniscus [45], bone TE [46], smooth muscle cell constructs for contractile muscle [47,48], and nerve regeneration [49][50][51], have been recently evaluated. For polyurethanes containing aromatic isocyanate moieties like methane-4,4'-diphenyldiisocyanate (MDI) and 2,4-toluenediisocyanate (TDI) toxic aromatic diamines were indicated after degradation.…”