Light Cross-Linkable Marine Collagen for Coaxial Printing of a 3D Model of Neuromuscular Junction Formation

Abstract

Collagen is a major component of the extracellular matrix (ECM) that modulates cell adhesion, growth, and migration, and has been utilised in tissue engineering applications. However, the common terrestrial sources of collagen carry the risk of zoonotic disease transmission and there are religious barriers to the use of bovine and porcine products in many cultures. Marine based collagens offer an attractive alternative and have so far been under-utilized for use as biomaterials for tissue engineering. Marine collagen can be extracted from fish waste products, therefore industry by-products offer an economical and environmentally sustainable source of collagen. In a handful of studies, marine collagen has successfully been methacrylated to form collagen methacrylate (ColMA). Our work included the extraction, characterization and methacrylation of Red Snapper collagen, optimisation of conditions for neural cell seeding and encapsulation using the unmodified collagen, thermally cross-linked, and the methacrylated collagen with UV-induced cross-linking. Finally, the 3D co-axial printing of neural and skeletal muscle cell cultures as a model for neuromuscular junction (NMJ) formation was investigated. Overall, the results of this study show great potential for a novel NMJ in vitro 3D bioprinted model that, with further development, could provide a low-cost, customizable, scalable and quick-to-print platform for drug screening and to study neuromuscular junction physiology and pathogenesis.

Directing Axonal Growth: A Review on the Fabrication of Fibrous Scaffolds That Promotes the Orientation of Axons

“…3D coaxial printing is another method used by researcher that combines a microfluidic approach with the exactness of computer aided scaffold fabrication [ 82 ]. In this particular study by Sanz, photolithography was also used to immediately solidify the scaffolds post fabrication [ 82 ]. Collagen methacrylate (ColMA) and gelatine methacrylate (GelMA) were used as bio-ink to produce a model of neuromuscular junction (NJM).…”

“…Collagen methacrylate (ColMA) and gelatine methacrylate (GelMA) were used as bio-ink to produce a model of neuromuscular junction (NJM). However, instead of mixing these hydrogels together, they were extruded from different nozzles to produce a construct made up of two separate materials that specializes in the culture of different cell types [ 82 ]. The methacrylation of collagen for immediate UV crosslinking after scaffolds fabrication improves its mechanical integrity.…”

Recent Applications of Biopolymers Derived from Fish Industry Waste in Food Packaging

“…Compared to mammalian collagen, marine collagen presents a comparable or slightly lower molecular weight and a lower denaturation (melting) temperature [ 25 ], which is about 20–35 °C for most fish species with higher values for collagen derived from warm–water species [ 53 , 69 ]. In order to enhance the thermal stability, suitable crosslinking treatments have been studied [ 70 , 71 ].…”

Tissue-Specific Decellularized Extracellular Matrix Bioinks for Musculoskeletal Tissue Regeneration and Modeling Using 3D Bioprinting Technology

“…Moreover, the integration of HNSCs facilitated rapid innervation, restoration, maturation, and functionalization of muscle tissue in a rodent model of muscle defect injury, suggesting that the neural-skeletal muscle construct can be rapidly integrated with the host neural network, leading to accelerated muscle regeneration. In addition, the coaxial 3D bioprinting technique has also been utilized to co-print primary myoblasts and NSC-34 cells (a motor neuron cell line) [144]. Such a core/shell structure mimics the muscle/neuron physiological system by bringing differentiating motor neuronlike cells in proximity with skeletal myoblasts.…”