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.
Highlights
Skeletal muscle tissues aid the body in support, locomotion, and even in the regulation of metabolism
Three-dimensional and functional muscle tissue constructs with optimized mechanical and biological properties are required in tissue regeneration and drug screening applications
The viscosity of gelatin methacryloyl (GelMA)-alginate hydrogels increased as the alginate concentration was increased (Figure 1b)
Summary
Skeletal muscle tissues aid the body in support, locomotion, and even in the regulation of metabolism. Bioprinting has allowed for the incorporation of cells and soluble factors during printing process [12] This process has recently become popular in fabricating different tissue and organ constructs due to its ability to mimic the hierarchical structure of the native tissues [13,14]. Testa et al used the same strategy and printed human muscle cells derived from perivascular and pericyte stem cells to treat sphincter muscle defects [18] These approaches used bioinks with limited tunability in physicochemical and biological properties. Alginate concentration within GelMA bioinks should be optimized to achieve the desired viscosity for bioprinting, and to obtain high cellular viability and function of skeletal muscles cells. By adding oxygen-releasing particles (i.e., calcium peroxide (CPO)) to the GelMA bioink, the optimal percentage of the CPO was used to enhance the viability and metabolic activity of C2C12 cells
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