Engineering muscle cell alignment through 3D bioprinting

Abstract

Processing of hydrogels represents a main challenge for the prospective application of additive manufacturing (AM) to soft tissue engineering. Furthermore, direct manufacturing of tissue precursors with a cell density similar to native tissues has the potential to overcome the extensive in vitro culture required for conventional cell-seeded scaffolds seeking to fabricate constructs with tailored structural and functional properties. In this work, we present a simple AM methodology that exploits the thermoresponsive behavior of a block copolymer (Pluronic ) as a means to obtain good shape retention at physiological conditions and to induce cellular alignment. Pluronic/alginate blends have been investigated as a model system for the processing of C2C12 murine myoblast cell line. Interestingly, C2C12 cell model demonstrated cell alignment along the deposition direction, potentially representing a new avenue to tailor the resulting cell histoarchitecture during AM process. Furthermore, the fabricated constructs exhibited high cell viability, as well as a significantly improved expression of myogenic genes vs. conventional 2D cultures. © 2017 Wiley Periodicals, Inc. J Biomed Mater Res Part A: 105A: 2582-2588, 2017.

The thickness of poly‐phenoxyethyl methacrylate brush interferes with cellular behavior and function of myofibers

“…Muscle tissue engineering and regeneration depends on the regenerative properties of the satellite cells and their potential for proliferation and differentiation . After harvesting from adult muscle and successfully grown in vitro , these satellite cells were activated as myoblasts and seeded into the natural (e.g., alginate, collagen, and fibrin) or synthetic (e.g., PGA, PLA, and PLGA) polymers, to try to organize functional muscle tissue . The PHEMA has been regarded as a potential scaffold material for muscle tissue engineering applications.…”

“…For engineering artificial skeletal muscle tissue in vitro , generate autologous satellite cells by biopsy, expand and differentiate cells in a 3D defined environment in an artificial bioreactor, and reimplant the neo‐tissue after differentiation has taken place . During this approach, 3D environment was fabricated by porous synthesized polymers, involving fibrin, PLG, polyglycolic acid (PGA), alginate, and others, and was taken as ECM composition to sustain the attachment, alignment, and differentiation of myoblasts. For promoting the survival and outward migration of myoblasts, the modified layer on substrate surface, including bioactive molecules and peptide motif, has been used to improve the synthesized polymers .…”

From Shape to Function: The Next Step in Bioprinting

“…Direct alignment of C2C12 myoblast cells along the main axis of bioprinted filaments has been observed and was attributed to the shear during extrusion. This led to maturation of confluent myotubes (enhanced myogenin, MyoD, and α‐sarcomeric actin mRNA levels) over 21 days of culture, a phenomenon that was not observed for cells maintained in 2D cultures . In another study using a fibrinogen–PEGDA semisynthetic hydrogel as bioink, confluent sarcomerogenesis and effective myoblastic maturation and stability was also obtained within two weeks.…”

Physiological culture conditions alter myotube morphology and responses to atrophy treatments: implications for in vitro research on muscle wasting

“…; Mozetic et al. ) biomolecular or inert supports, and use of electrical stimulation (Thelen et al. ; Bayol et al.…”

“…Several authors have tested a variety of approaches to improve myotube growth, differentiation and survival, with some success. These have included the use of two- (Langen et al 2003;Cooper et al 2004;DeQuach et al 2010) or three-dimensional (Kroehne et al 2008;Mozetic et al 2017) biomolecular or inert supports, and use of electrical stimulation (Thelen et al 1997;Bayol et al 2005;Boonen et al 2011;van der Schaft et al 2013). However, such experimental conditions have not been widely adopted by other research groups, nor have there been attempts to test myotube culture conditions that combine several features of the in vivo milieu to study disease-related muscle atrophy.…”