Novel ultrashort self-assembling peptide bioinks for 3D culture of muscle myoblast cells

Abstract

The ability of skeletal muscle to self-repair after a traumatic injury, tumor ablation, or muscular disease is slow and limited, and the capacity of skeletal muscle to self-regenerate declines steeply with age. Tissue engineering of functional skeletal muscle using 3D bioprinting technology is promising for creating tissue constructs that repair and promote regeneration of damaged tissue. Hydrogel scaffolds used as biomaterials for skeletal muscle tissue engineering can provide chemical, physical and mechanical cues to the cells in three dimensions thus promoting regeneration. Herein, we have developed two synthetically designed novel tetramer peptide biomaterials. These peptides are self-assembling into a nanofibrous 3D network, entrapping 99.9% water and mimicking the native collagen of an extracellular matrix. Different biocompatibility assays including MTT, 3D cell viability assay, cytotoxicity assay and live-dead assay confirm the biocompatibility of these peptide hydrogels for mouse myoblast cells (C2C12). Immunofluorescence analysis of cell-laden hydrogels revealed that the proliferation of C2C12 cells was well-aligned in the peptide hydrogels compared to the alginate-gelatin control. These results indicate that these peptide hydrogels are suitable for skeletal muscle tissue engineering. Finally, we tested the printability of the peptide bioinks using a commercially available 3D bioprinter. The ability to print these hydrogels will enable future development of 3D bioprinted scaffolds containing skeletal muscle myoblasts for tissue engineering applications.

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

“…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 ].…”

“…To overcome this shortcoming, Arab et al. used printable, self-assembling peptides to create 3D hydrogel constructs to promote myogenic differentiation [ 87 ]. The authors found that cells within their peptide hydrogels aligned in response to the structural cues provided by their synthetic peptides, while cells grew in a random orientation in their alginate-gelatin controls.…”

Applications of self-assembling ultrashort peptides in bionanotechnology

“…Recently, Arab et al used two tetra-peptides, CH-01 and CH-02, as bioinks for 3D culture of C2C12 mouse myoblasts. 71…”

Optimization of a 3D bioprinting process using ultrashort peptide bioinks

“…Due to their amphiphilic character and their innate tendency to self-assemble in water, they form rapidly nanofibrous scaffolds in an aqueous solution in forms of soft solid and transparent hydrogels. The natural but synthetic character of these self-assembling peptides renders them as appealing bioinks for bioprinting[ 8 ]. Peptides are generally known for their biocompatibility, biodegradability, and suitability for cell growth[ 9 ].…”