Composite Biomaterials as Long‐Lasting Scaffolds for 3D Bioprinting of Highly Aligned Muscle Tissue

Abstract

New biocompatible materials have enabled the direct 3D printing of complex functional living tissues, such as skeletal and cardiac muscle. Gelatinmethacryloyl (GelMA) is a photopolymerizable hydrogel composed of natural gelatin functionalized with methacrylic anhydride. However, it is difficult to obtain a single hydrogel that meets all the desirable properties for tissue engineering. In particular, GelMA hydrogels lack versatility in their mechanical properties and lasting 3D structures. In this work, a library of composite biomaterials to obtain versatile, lasting, and mechanically tunable scaffolds are presented. Two polysaccharides, alginate and carboxymethyl cellulose chemically functionalized with methacrylic anhydride, and a synthetic material, such as poly(ethylene glycol) diacrylate are combined with GelMA to obtain photopolymerizable hydrogel blends. Physical properties of the obtained composite hydrogels are screened and optimized for the growth and development of skeletal muscle fibers from C2C12 murine cells, and compared with pristine GelMA. All these composites show high resistance to degradation maintaining the 3D structure with high fidelity over several weeks. Altogether, in this study a library of biocompatible novel and totally versatile composite biomaterials are developed and characterized, with tunable mechanical properties that give structure and support myotube formation and alignment.

3D Bioprinting in Skeletal Muscle Tissue Engineering

“…Poly(vinyl alcohol)(PVA)‐alginate has also been used to bioprint constructs with micropores showing controlled release of proteins . In another study, Ramon‐Azcon and colleagues developed a library of composite biomaterials used as bioinks and showed that GelMA‐carboxymethyl cellulose methacrylate (CMCMA) allowed the growth and differentiation of encapsulated C2C12, while presenting high resistance to degradation . Moreover, microfluidic print heads have also been used to deposit multiple materials into microfibers or droplets.…”

“…[90] In another study, Ramon-Azcon and colleagues developed a library of composite biomaterials used as bioinks and showed that GelMA-carboxymethyl cellulose methacrylate (CMCMA) allowed the growth and differentiation of encapsulated C2C12, while presenting high resistance to degradation. [91] Moreover, microfluidic print heads have also been used to deposit multiple materials into microfibers or droplets. This technique allows fast switching between different bioinks as well as the formation of fibers with different coded bioink segments or heterogeneous fibers with two or more parallel bioinks in the fiber.…”

A three-dimensional bioprinted model to evaluate the effect of stiffness on neuroblastoma cell cluster dynamics and behavior

“…Methacrylated gelatin (GelMA), a photocrosslinkable hydrogel derived from natural gelatin, emulates the ECM for various cells types in combination with non-biodegradable materials such as alginate. We used the synthesis process as previously described 43 . Briefly, Gelatin (Sigma-Aldrich, USA) was modified to a 40% degree of methacrylation.…”

Engineered 3D Polymer and Hydrogel Microenvironments for Cell Culture Applications

“…The alginate/nano-cellulose bioink system, including variations such as sodium alginate [268], alginate sulphate [267], and the various phases of nano-cellulose, have been reported to have better performance than, for example, hyaluronic acid/nano-cellulose [266]. Gelatin/nano-cellulose based bioinks combined with oxidation also have no cytotoxicity, a pore size of~600 µm, good cell viability, improved mechanical properties (compressive modulus from 0.5 to 8.5 kPa with the addition of 2% w/v nano-cellulose), and improved print properties [284][285][286]. PEGDA/nano-cellulose structures fabricated by SLA showed high NIH 3T3 cell viability [274].…”