Three-Dimensional Bioprinting of Functional Skeletal Muscle Tissue Using GelatinMethacryloyl-Alginate Bioinks

Abstract

Skeletal muscle tissue engineering aims to fabricate tissue constructs to replace or restore diseased or injured skeletal muscle tissues in the body. Several biomaterials and microscale technologies have been used in muscle tissue engineering. However, it is still challenging to mimic the function and structure of the native muscle tissues. Three-dimensional (3D) bioprinting is a powerful tool to mimic the hierarchical structure of native tissues. Here, 3D bioprinting was used to fabricate tissue constructs using gelatin methacryloyl (GelMA)-alginate bioinks. Mechanical and rheological properties of GelMA-alginate hydrogels were characterized. C2C12 myoblasts at the density 8 × 106 cells/mL were used as the cell model. The effects of alginate concentration (0, 6, and 8% (w/v)) and crosslinking mechanism (UV crosslinking or ionic crosslinking with UV crosslinking) on printability, cell viability, proliferation, and differentiation of bioinks were studied. The results showed that 10% (w/v) GelMA-8% (w/v) alginate crosslinked using UV light and 0.1 M CaCl2 provided the optimum niche to induce muscle tissue formation compared to other hydrogel compositions. Furthermore, metabolic activity of cells in GelMA bioinks was improved by addition of oxygen-generating particles to the bioinks. It is hoped that such bioprinted muscle tissues may find wide applications in drug screening and tissue regeneration.

Biofabrication of muscle fibers enhanced with plant viral nanoparticles using surface chaotic flows

“…Figure 3B shows the PrestoBlue ® assay results, conducted to measure the metabolic activity of our cell-seeded fibers. We observed a gradual and substantial increase in activity over the first 12 days in all cases (i.e., GelMA fibers with or without TuMV), which is expected at proliferation stages and has also been reported for C2C12 cells seeded on GelMA scaffolds [36,37]. All samples followed approximately the same trend the first 6 days, but in the following days they exhibited significantly different behaviors (p*<0.05).…”

Advances in biomaterials for skeletal muscle engineering and obstacles still to overcome

“…However, recent research has focused on developing open-source software or code to more easily and accurately quantify myotube formation parameters without incorporating human error or bias [ 102 ]. Other myogenic markers that are commonly used to visualize and assess myotube formation, include desmin [ 82 , 103 ] and α-sarcomeric actin [ 82 , 85 , 88 , 89 , 101 ]. In addition, a cytoskeletal actin stain can be utilized to assess overall biomaterial-guided cell alignment [ 50 , 70 , 72 , 76 , 84 , 87 , 98 ].…”

Recent progress in extrusion 3D bioprinting of hydrogel biomaterials for tissue regeneration: a comprehensive review with focus on advanced fabrication techniques

“…Mimicking extremely packed and arranged cellular structure of the native muscle tissue, employing natural or synthetic scaffolds and microscale technologies, is essential for the successful SMTE [270,271]. The 3D bioprinting has emerged as a powerful microscale technology for SMTE [272,273].…”

“…Different studies have investigated the propriety of GelMA hydrogel and its composites with various nanomaterials for SMTE [72,265,279,280]. Some solutions have been proposed to provide high cellular viability and function of skeletal muscle cells, such as applying optimized alginate concentration combined with a suitable crosslinking method [272,281]. The administration of growth factors (locally or systemically) has also presented a great promise to stimulate angiogenesis, stem cell recruitment and differentiation, cell survival and proliferation, a decrease of apoptosis, and adaptive remodeling [282,283].…”